EP0450060A1 - Sequen age d'adn - Google Patents

Sequen age d'adn

Info

Publication number
EP0450060A1
EP0450060A1 EP19910900474 EP91900474A EP0450060A1 EP 0450060 A1 EP0450060 A1 EP 0450060A1 EP 19910900474 EP19910900474 EP 19910900474 EP 91900474 A EP91900474 A EP 91900474A EP 0450060 A1 EP0450060 A1 EP 0450060A1
Authority
EP
European Patent Office
Prior art keywords
dna molecule
complementary dna
incorporated
subject
molecule
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP19910900474
Other languages
German (de)
English (en)
Inventor
Roger Y. Tsien
Pepi Ross
Margaret Fahnestock
Allan J. Johnston
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
SRI International Inc
Original Assignee
SRI International Inc
Stanford Research Institute
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by SRI International Inc, Stanford Research Institute filed Critical SRI International Inc
Publication of EP0450060A1 publication Critical patent/EP0450060A1/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6869—Methods for sequencing
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor

Definitions

  • This invention relates to DNA sequencing. More particularly, it relates to methods and apparatus for determining the sequence of deoxyribonucleotides within DNA molecules .
  • DNA sequencing is an important tool.
  • a current goal of the biological community in general is the determination of the complete structure of the DNA of a number of organisms, including man. This information will aid in the understanding, diagnosis, prevention and treatment of disease.
  • the DNA to be sequenced is enzymatically copied by the Klenow fragment of DNA polymerase I or by a similar polymerase enzyme such as Taq polymerase or SequenaseTM .
  • the enzymatic copying is carried out in quadruplicate.
  • a low concentration of a chain terminating dideoxynucleotide is present, a different dideoxynucleotide being present in each of the four reactions (ddATP, ddCTP, ddGTP and ddTTP) .
  • the polymerase reaction is terminated, again producing sets of nested fragments. Again, the nested fragments have to be separated from one another by electrophoresis to determine the sequence.
  • the present invention provides methods and apparatus for determining the sequence of deoxyribonucleotides in a DNA molecule.
  • a key characteristic of this invention is that it determines the DNA sequence without recourse to electrophoresis or other size-based separation techniques.
  • the present invention provides a method for determining the deoxyribonucleotide sequence, of a single stranded DNA subject molecule.
  • This method involves synthesizing, in the presence of a multitude of identical copies of the subject DNA, the DNA molecule which is complementary to it.
  • This synthesis is carried out using deoxyribonucleotide triphosphates (dNTP) in a stepwise serial manner so as to simultaneously build Up numerous copies of the complementary molecule, dNTP by dNTP.
  • dNTP deoxyribonucleotide triphosphates
  • this invention provides apparatus for carrying out the above-described method.
  • this method and apparatus for carrying it out can take many different configurations.
  • This invention can be carried out in a single reaction zone with multiple differentiable reporters or in multiple reaction zones with a single reporter in each zone. It can be carried out by detecting the incremental signal change after addition of reporters or by noting each added reporter separately. The various reporters can be measured in the reaction zones while attached to the growing molecule or they can be separated from the molecule and then measured.
  • the invention can be practiced to create the growing complementary DNA chain without interruption or it can be practiced in stages wherein a portion of the complementary chain is created and its sequence determined; this portion of the chain is then removed; a sequence corresponding to a region of the removed chain is separately synthesized and used to prime the template chain for subsequent chain growth. The latter method can be repeated as needed to grow out in portions the complete complementary chain.
  • Figures 1A and IB are schematic diagrams of the process of this invention on a molecular level.
  • Figure 2 is a schematic representation of one form of apparatus for practising the invention.
  • the DNA growth takes place in a single reaction zone.
  • This embodiment uses separate, distinguishable reporters associated with each of the four nucleotides incorporated into the growing molecule. The four different reporters are measured after each addition to detect which base has just been added to that position of the complementary chain.
  • Figure 3 is a schematic representation of another form of apparatus for practising the invention.
  • This embodiment employs four reaction zones in which the molecular growth is carried out in quadruplicate. In each of the four zones, a different one of the four nucleotides is associated with a reporter (with the remaining three being unlabeled) so that the identity of the nucleotide incorporated at each stage can be determined.
  • Figure 4 is a schematic representation of an adoption of the apparatus for practising the invention particularly adapted for carrying out the invention to grow a series of portions of the complementary molecule as opposed to a single continuous complementary molecule.
  • Figures 5 through 8 are pictorial representations of chemical reaction sequences which can be used to synthesize representative labeled nucleotide building blocks for use in the practice of this invention.
  • dNTPs of these materials are abbreviated as dATP, dCTP, dGTP and dTTP. When these materials are blocked in their 3'-OH position they are shown as 3 'blockeddATP, 3'blockeddCTP, 3 'blockeddGTP and 3 'bl ⁇ ckeddTTP. Similarly, when they are each tagged or labeled with a common reporter group, such as a single fluorescent group, they are represented as dA'TP, dC'TP, dG'TP and dT'TP.
  • a solid support 1 is illustrated with a reactive group A attached to its surface via tether 2. This attachment can be covalent, ionic or the like.
  • a second reactive group. X capable of bonding to group A, again via a covalent, ionic or the like bond, is attached to the 5' end of a DNA primer 4.
  • This primer has a known DNA sequence. When coupled to the substrate via the A-X bond it forms immobilized primer 5.
  • Primer 5 is then hybridized to template DNA strand 6 which is made up of an unknown region 7 inserted between regions 8 and 8 ' . Regions 8 and 8' are located at the 5' and 3' ends of the unknown region and have known sequences.
  • the 8' region's known sequence is complementary to the sequence of primer 4 so that those regions hybridize to form immobilized template DNA 9. Therefore the individual dNTPs are serially added to form the DNA sequence complementary to the unknown region of the template. 11 and 12 represent the first two such dNTPs incorporated into the growing molecule. These in turn provide the identity of their complements 11' and 12' respectively. This growth continues until the entire complementary DNA molecule has been constructed. Completion can be noted by identifying the sequence corresponding to the 8 region of template 6. Turning to Figure IB, a variation of this chemistry is shown in that the template 6* carries the reactive group X which bonds to the substrate via the A-X bond to form an immobilized template 5*.
  • Device 13 for carrying out the invention is shown schematically.
  • Device 13 includes a reaction zone 14 which carries inside it a surface 15.
  • a plurality of copies of a subject primed single stranded DNA are immobilized on this surface 15. This is the strand of DNA for which the sequence is desired.
  • the immobilized DNA is depicted fancifully on surface 15 as if it were present as a series of separately visible attached strands. As will be appreciated, this is not in fact the case and is only done to guide the reader as to the location of the DNA strands .
  • the reaction zone 14 may be configured to permit direct reading of reporter signals emanating from within. Examples of this configuration include equipping the reaction zone to permit measuring fluorescence or luminescence through one or more transparent walls or detecting radionuclide decay.
  • Reaction zone 14 is fitted with inlet 16 for the addition of polymerase or another suitable enzyme capable of moderating the templat ⁇ e-directing coupling of nucleotides to one another.
  • the reaction zone is ⁇ also accessed by inlet lines, 18a-18d for four differently labeled blocked dNTPs, that is 3'blockeddA'TP, 3 'blockeddC' 'TP, 3 'blockeddG' ' 'TP, and 3'blockeddT' ' ' 'TP. These materials can be added in four separate lines, as shown, or can be premixed, if desired, and added via a single line. Buffer and other suitable reaction medium components are added via line 20.
  • the polymerase and the four labeled dNTPs are added to the reaction zone 14 under conditions adequate to permit the enzyme to bring about addition of the one, and only the one, of the four labeled blocked dNTPs which is complementary to the first available template nucleotide following the primer.
  • the blocking group present on the 3 '-hydroxyl position of the added dNTP prevents inadvertent multiple additions .
  • the liquid in reaction zone 14 is drained through line 22 either to waste, or if desired to storage for reuse.
  • the reaction zone and the surface 15 are rinsed as appropriate to remove unreacted, uncoupled labeled blocked dNTPs.
  • the first member of the complementary chain is now in place associated with the subject chain attached to surface 15.
  • the identity of this first nucleotide can be determined by detecting and identifying the label attached to it.
  • This detection and identification can be carried out in the case of a fluorescent label by irradiating the surface with a fluorescence-exciting beam from light source 24 and detecting the resulting fluorescence with detector 26.
  • the detected florescence is then correlated to the fluorescence properties of the four different labels present on the four different deoxynucleotide triphosphates to identify exactly which one of the four materials was incorporated at the first position of the complementary chain. This identity is then noted.
  • a reaction is carried out to remove the blocking group and label from the 3' position on the first deoxynucleotide triphosphate.
  • This reaction is carried out in reaction zone 14.
  • a deblocking solution is added via line 28 to remove the 3' hydroxyl labeled blocking group. This then generates an active 3' hydroxyl position on the first nucleotide present in the complementary chain and makes it available for coupling to the 5' position of the second nucleotide.
  • removal of the deblocking solution via line 22 and rinsing as needed the four blocked, labeled deoxynucleotide triphosphates, buffer and polymerase are again added and the appropriate second member is then coupled into the growing complementary chain.
  • the second member of the chain can be identified based on its label. This process is then repeated as needed until the complementary chain has been completed.
  • the sequence of incorporated deoxynucleotides is known, and therefore so is the sequence of the complement which is the subject chain. It will be appreciated that this process is easily automated. It is a series of fluid additions and removals from a reaction zone. This can be easily accomplished by a series of timer-controlled valves and the like. This technology has been well developed in the area of oligonucleotide synthesizers, peptide synthesizers, and the like. In such an automated system, the timing can be controlled by a microprocessor or, in most cases, by a simple programmable timer. The rate and " extent of reaction can be monitored by measurement of the reporter concentration at various stages .
  • the labels present in the blocked dNTPs can be incorporated in one of several manners. For one, they can be incorporated directly and irremovably in the deoxynucleotide triphosphate unit itself. Thus, as the complementary chain grows there is a summing of signals and one identifies each added nucleotide by noting the change in signal observed after each nucleotide is added.
  • the label is incorporated within the blocking group or is otherwise incorporated in a way which allows it to be removed between each addition. This permits the detection to be substantially simpler in that one is noting the presence of one of the four reporter groups after each addition rather than a change in the sum of a group of reporter groups .
  • the presence of reporter signal is noted directly in the reaction zone 14 by the analytical system noted as source 24 and detector 26. It will be appreciated, however, that in embodiments where the reporter group is removed during each cycle, it is possible to read or detect the reporter at a remote site after it has been carried out of the reaction zone 14.
  • drain line 22 could be valved to a sample collector (not shown) which would isolate and store the individual delabeling product solutions for subsequent reading.
  • the various removed labels could be read as they flowed out of the reaction zone by equipping line 22 with an in-line measurement cell such as source 24' and detector 26' or the like.
  • a second embodiment of this invention employs four separate parallel reaction zones. This method has the advantage of requiring only one type of labeling and being able to use it with all four dNTPs.
  • Figure 3 shows a schematic representation of a device 30 which has the four reaction zone configuration. In this configuration there are four reaction zones 32a through 32d, each of which resembles the reaction zone 14 in Figure 2. In these cases each of the four reaction zones contains a surface 34a-d to which is immobilized numerous copies of a primed subject single stranded DNA. Each reaction zone is supplied with polymerase via lines 36a-d. Each zone is supplied with suitable reaction medium via lines 38a-38d. The four dNTPs are supplied in blocked form to each zone, as well.
  • zone 32a one of the blocked dNTPs is labeled, for example "A'"; in zone 32b a second dNTP is labeled, for example "C”; in zone 32c a third dNTP is labeled, for example "G'”; and in 32d the fourth labeled dNTP "T' M is present.
  • These labeled materials are supplied via lines 40a through 40d respectively.
  • Unlabeled blocked dNTPs are supplied via lines 42a-d so that each of the four reaction zones contains three unlabeled blocked dNTPs and one labeled blocked dNTP.
  • the various labeled and unlabeled dNTP ' s can be premixed. These premixed materials can be added to the various reaction zones via single addition lines.
  • the single stranded DNA hybridized to a primer and attached to each of surfaces 34a-34d is contacted with polymerase (supplied via lines 36a-36d), buffer (supplied via lines 38a-38d) and the four bases in each of the four reaction zones .
  • the blocked dNTP which complements the first base on the subject chain couples.
  • this base is labeled.
  • this label is incorporated into the growing chain, one can determine the identity of the dNTP which is incorporated at the first position. This determination of the identity of the first unit of the chain can be carried out using signal sources and detectors such as 44a-44d and 46a-46d, respectively.
  • Deblocking is carried out by adding deblocking solution to the reaction zone through lines 48a-48d.
  • Lines 50a-50d are drain lines for removing material from the reaction zones following each step.
  • all of the variations noted with reference to the device described in Figure 2 can also be used including cumulating reporter signals and generating reporter signals away from the reaction zone by removing the reporter groups as part of each of the sequential couplings.
  • this embodiment can be readily automated, as well.
  • One obvious potential shortcoming of the present invention is that it employs a long sequence of serial reactions. Even if the efficiency and yield of each of these reactions are relatively high, the overall yield becomes the product of a large number of numbers, each of which is somewhat less than 1.00, and thus can become unacceptably low. For example if the yield of a given addition step is 98% and the deblocking is 98% as well, the overall yield after 15 additions is 48 , after 30 additions it is 23% and after 60 additions it is 5.3%.
  • This limitation can be alleviated by periodically halting the DNA molecule growth and using the sequence data obtained prior to halting the growth to externally recreate a portion of the molecule which can then be used as a primer for renewed DNA fabrication. This process is illustrated in Figure 4.
  • FIG 4 shows a schematic of an automated sequencer 52 employing the present invention.
  • Sequencer 52 has a single reaction zone 14 combining the subject primed DNA, immobilized therein such as on surface 15.
  • the four 3-blocked DNTP's are fed to the reaction zone through line 18.
  • Polymerase and buffers are added via lines 16 and 20, respectively.
  • the dNTP 's, polymerase and buffer can be recycled from step to step via lines 54 and 56 and holding vessel 58. All of the valves admitting and removing fluids from reaction zone 14 can be controlled by central computer 60 which functions as a valve control clock.
  • This computer 60 can also control the addition of deblocker from line 28, deblocking eluent with cleaved labels (as obtained when the label is present in the blocking group) is removed via line 22 and detected via detector system 24/26 reading label values in detector vessel 62.
  • This embodiment illustrates the use of a fluorescent label system and shows the addition of fluorescent sensitizer (flooder) via line 64 to the fluorescent detection zone 62.
  • the deblocking solution and detected label are discarded via line 66.
  • the signal presented by the label identified by detector 26 is passed to analog/digital converter 68 and therein to a memory in central computer 60 where it is stored.
  • the memory in computer 60 contains the sequence of an initial portion of the complementary DNA molecule which has been constructed in association with the subject or target DNA molecule contained within reactor 14. After some number of units have been assembled - typically 25 to 300, or more; preferably 50 to 300, or more; and more preferably 100 to 300, or more - the growing complementary DNA molecule is stripped from the immobilized subject DNA molecule and discarded. This stripping (denaturing) can be done by art-known methods such as by warming the reaction zone to 75°C or higher (preferably 90-95°C) for a few (1-15) minutes.
  • the sequence information stored in computer 60 is used to drive DNA synthesizer 70 to externally create a new DNA primer corresponding to at least a portion of the discarded DNA molecule. (The sequence can also be read on printer 72, if desired.)
  • This newly constructed DNA primer molecule is fed through line 74 to reaction zone 14 under hybridization conditions so as to join to the complementary region of the subject DNA molecule as a new primer.
  • the length of the primer must be adequate to. unambiguously and strongly hybridize with a single region of the subject DNA. As is known in the hybridization art, this can depend upon factors such as the sequence, environmental conditions, and the length of the subject DNA. For efficiency of operation, the primer should ideally be as short as possible.
  • Primer lengths typically range from about 10 bases to about 30 bases, although shorter primers would certainly be attractive if they met the above criteria, and longer primers could be used albeit with an increase in cost and time. Good results generally are achieved with primers from 12 to 20 bases long. This gives the molecular growth reaction a "new start" with a large number of properly primed identical molecules. This allows a strong signal to be generated when the next dNTP is coupled.
  • the coupling process employed in this invention to incorporate each of the blocked deoxynucleotide triphosphates into the growing complementary chain is an enzyme moderated process.
  • Each member of the complementary DNA chain is added using a suitable template-dependent enzyme.
  • One enzyme which can be used is Sequenase TM enzyme (an enzyme derived from bacteriophage 7 DNA polymerase that is modified to improve its sequencing properties - see Tabor and
  • Sequenase TM examples include but are not limited to
  • the coupling conditions which are employed are those known in the art for these enzymes.
  • these include temperatures in the range of from about room temperature to about 45 C; a buffer of pH 7 to 8 and preferably pH 7.3 to 7.7; an enzyme concentration of from about 0.01 units per microliter to about 1 unit per microliter and a reaction time of from about 1 to about 20 minutes and preferable 1 to 5 minutes.
  • a typical buffer for use with Sequenase TM is made up of
  • these typical conditions include temperatures in the range of from about 10 C to about 45 C and preferably from about 15°C to about 40°C; a buffer of pH 6.8 to 7.4 and preferably pH 7.0 to 7.4; an enzyme concentration of from about 0.01 units per microliter to about 1 unit per microliter and preferably from about 0.02 to about 0.15 units per microliter and a reaction time of from about 1 to about 40 minutes.
  • a typical buffer for use with Klenow fragment of DNA polymerase I is made up of
  • 3 '-blocking groups include: (l) the ability of a polymerase enzyme to accurately and efficiently incorporate the dNTPs carrying the 3 '-blocking groups into the cDNA chain,
  • the 3 '-blocking group carries a reporter group, it is desirable that the reporter permit sensitive detection either when part of the cDNA chain before deblocking or subsequent to deblocking in the reaction eluant.
  • 3 '-blocked dNTPs are used that can be incorporated in a template-dependent fashion and easily deblocked to yield a viable 3 ' -OH terminus.
  • the most common 3 '-hydroxyl blocking groups are esters and ethers.
  • ester blocking groups such as lower (1-4 carbon) alkanoic acid and substituted lower alkanoic acid esters, for example formyl, acetyl, isopropanoyl, alpha fluoro- and alpha chloroacetyl esters and the like; ether blocking groups such as alkyl ethers; phosphate blocking groups; carbonate blocking groups such as 2-nitrobenzyl; 2,4-dinitrobenzene-sulfenyl and tetrahydrothiofuranyl ether blocking groups.
  • Blocking groups can be modified to incorporate reporter moieties, if desired, including radiolabels (tritium, C 14 or F ⁇ ** 2 , for example), enzymes, fluorophores and chromophores .
  • selectively-removable amine protection groups include carbamate ⁇ cleavable by acid hydrolysis [t-butyl, 2-(biphenyl)isopropyl] and certain amides susceptible to acid cleavage (formamide, trichloroacetamide) (Greene, 1981) .
  • nucleotide derivatives protection of the primary amino groups is performed prior to phosphonation.
  • standard amino protecting groups cleavable by ammonolysis may be used.
  • the sequencing scheme After successfully incorporating a 3 '-blocked nucleotide into the DNA chain, the sequencing scheme requires the blocking group to be removed to yield a viable 3 '-OH site for continued chain synthesis.
  • the deblocking method should:
  • the exact deblocking chemistry selected will, of course, depend to a large extent upon the blocking group employed. For example, removal of ester blocking groups from the 3 'hydroxyl function is usually achieved by base hydrolysis. The ease of removal varies widely; generally, the greater the electro-negativity of substituents on the carbonyl carbon, the greater the ease of removal. For example, the highly electronegative group trifluoroacetate is cleaved rapidly from 3' hydroxyls in methanol at pH 7 (Cramer et al. , 1963) and thus would not be stable during coupling at that pH.
  • Phenoxyacetate groups are cleaved in less than one minute but require substantially higher pH such as is achieved with NH-/ methanol (Reese and Steward, 1968).
  • the ester deblocking rate is advantageously selected so as to exhibit a deblocking rate of less than 10 -3s-1 during the incorporation, and at least 10 ⁇ s during the deblocking stage. Ideally, this rate change is achieved by changing the buffer pH from 7 to about 10, but care must be taken not to denature the DNA.
  • hydroxyl blocking groups are cleaved selectively using chemical procedures other than base hydrolysis.
  • 2,4-Dinitrobenzenesulfenyl groups are cleaved rapidly by treatment with nucleophiles such as thiophenol and thiosulfate (Letsinger et al., 1964).
  • Allyl ethers are cleaved by treatment with Hg(II) in acetone/water (Gigg and Warren, 1968) .
  • Tetrahydrothiofuranyl ethers are removed under neutral conditions using Ag(I) or Hg(II) (Cohen and Steele, 1966; Cruse et al . , 1978).
  • These protecting groups which are stable to the conditions used in the synthesis of dNTP analogues and in the sequence incorporation steps, have some advantages over groups cleavable by base hydrolysis - deblocking occurs only when the specific deblocking reagent is present and premature deblocking during incorporation is minimized.
  • Photochemical deblocking can be used with photochemically-cleavable blocking groups.
  • Several blocking groups are available for such an approach.
  • the use of o-nitrobenzylethers as protecting groups for 2 '-hydroxyl functions of ribonucleosides is known and demonstrated (Ohtsuka et al. , 1978); removal occurs by irradiation at 260 nm.
  • Alkyl o-nitrobenzyl carbonate protecting groups are also cleaved by irradiation at pH 7 (Cama and Christensen, 1978).
  • Enzymatic deblocking of 3 '-OH blocking groups is also possible. It has been demonstrated that T4 polynucleotide kinase can convert 3 '-phosphate termini to 3 '-hydroxyl termini that can then serve as primers for DNA polymerase I (Henner et al . , 1983). This 3 '-phosphatase activity is used to remove the 3 '-blocking group of those dNTP analogues that contain a phosphate as the blocking group; the radioactive label enables the incorporation of the nucleotide analogue and the removal of the phosphate group to be followed easily. . If the use of radioisotopes represents too great a drawback, it is possible to use unlabeled phosphate monoesters with a cleavable fluorescent label (see below).
  • each dNTP into the complementary chain is noted by detecting a label or reporter group present in or associated with the incorporated dNTP.
  • the labels or markers are "innocuous".
  • An "innocuous marker or label or reporter” refers to a radioactive, fluorescent, or the like marker or reporter which has physical and chemical properties which do not interfere with either the enzymatic addition of the marked nucleotide to the cDNA, or the subsequent deblocking to yield a viable 3 '-OH terminus .
  • One simple labeling approach is to incorporate a radioactive species within the blocking group or in some other location of the dNTP units. This can be done easily by C 14 labeling or P32 labeling.
  • Another labeling approach employs fluorescent labels. These can be attached to the dNTP's via the 3 '0H- blocking groups or attached in other positions. There are two general routes available using fluorescent tags:
  • the first route is fairly straightforward and can employ a range of known fluorophores such as rhodamines, fluoresceins and the like, typically including those fluorophores known as useful in labeling dNTP's and the like.
  • fluorophores such as rhodamines, fluoresceins and the like, typically including those fluorophores known as useful in labeling dNTP's and the like.
  • the second route can employ a fluorophore where only a fragment is attached to the dNTP. This can reduce size and minimize steric interference. In the second route, rapid reaction of a normally nonfluorescent probe or molecule with specific functional group(s) found only on the label fragment leads to the formation of a fluorescent addition product. This leads to a signal only when the particular label
  • Blocking groups or other label fragment groups containing free thiol functions can be used for this approach.
  • the blocking group or other label fragment can contain a metal-binding ligand, e.g. a carboxylic acid group which will react with added rare earth metal ions such as europium or terbium ions to yield a fluorescent species.
  • This dNTP can be incorporated and the fluorescence measured and removed according to the methods described below.
  • One method involves the use of a fluorescent tag attached to the base moiety.
  • the tag may be chemically cleaved (either separately from or simultaneously with the deblocking step) and measured either in the reaction zone before deblocking or in the reaction eluant after cleavage.
  • the fluorescent moiety or other innocuous label can be attached to the dNTP through a spacer or tether.
  • the tether can be cleavable if desired to release the fluorophore or other label on demand.
  • Typical tethers are from about 2 to about 20, and preferably from about 3 to about 10 atoms in length.
  • the C-8 position of the purine structure presents an ideal position for attachment of a label.
  • Sarfati et al . (1987) describes a derivatization of deoxyadenosine at C-8 of the purine to prepare, ultimately, an 8-substituted biotin aldylamino dATP.
  • the Sarfati et al . (1987) approach can be used to prepare the appropriate fluorescent, rather than biotinylated, analogues.
  • a number of approaches are possible to produce fluorescent derivatives of thymidine and deoxycytidine.
  • One quite versatile scheme is based on an approach used by Prober et al . (1987) to prepare ddNTPs with fluorescent tags. Structures A, B, C and D below illustrate the type of fluorescent dNTPs that result from these synthetic approaches.
  • the synthetic routes have a great flexibility in that the linker can be varied with respect to length or functionality.
  • the terminal fluorescent moiety can also be varied according to need.
  • the labels so incorporated in the growing cDNA chain are detected by conventional analytical methods .
  • increased detection sensitivity is a major advantage of the present method.
  • the signal is based on a low level of fluorophores and is superimposed on a background of scatter from the gel and glass plates. This decreases sensitivity and often constrains current methods to the use of laser illumination to maximize sensitivity (Smith et al., 1986; Prober et al . , 1987; Ansorge et al . , 1986) .
  • Detection of fluorophores is readily achievable in commercial non-excited spectrofluorometers , such as are sold by Perkin-Elmer.
  • LED light- emitting diodes
  • Typical LEDs include:
  • Red LED emitting at approximately 650 nm
  • Green LED emitting at approximately 540 nm
  • Blue LED emitting at approximately 450 nm
  • the solution containing cleaved blocking groups or nucleotides is directly injected into a field ionization mass spectrometer. Identification of the particular nucleotide incorporated or cleaved is achieved by monitoring the relative abundance of molecular ion peaks corresponding to the specific nucleotides or blocking groups; for example, four distinct acetyl blocking groups differing by one mass unit (replacement of 0 to 3 hydrogens by deuterium) could be detected by monitoring a small “window. "
  • Immobilization of Subject DNA In the present invention, single stranded subject DNA or its primer is immobilized.
  • One approach to this immobilization is to attach the DNA to a solid substrate.
  • DNA and RNA are commonly attached noncovalently through ionic interactions along their length to various types of membranes (Southern, 1975; Maniatis, Fritsch, and Sambrook, 1982; Chuvpilp and Kravchenko, 1984).
  • polynucleotides are covalently attached along their length to membranes (Goldberg, et al . , 1979), resins (Seed, 1982; Arndt-Jovin, et al .
  • the inner quartz or glass surface can be advantageously functionalized using silanizing reagents such as triethoxysilylpropylamine or dichlorodi ethylsilane. This is followed by covalent attachment of a long-chain alkylamine to these functionalizing groups.
  • the single stranded subject DNA is attached to the long chain amine.
  • immobilization is carried out by attaching the subject DNA to a plastic surface.
  • a thin polypropylene chamber wall designed to pass Cer'enkov radiation from 32P, for example, can serve as a suitable substrate for DNA immobilization.
  • a plastic surface it is preferable to use the method of Kremsky et al .
  • the reaction zone has one or more openings covered with a membrane such as an ultrafiltration membrane, for example, Amicon's PM-5 or PM-10 membranes which have nominal molecular weight cut offs of 5000 and 10,000 respectively.
  • a membrane such as an ultrafiltration membrane, for example, Amicon's PM-5 or PM-10 membranes which have nominal molecular weight cut offs of 5000 and 10,000 respectively.
  • the single stranded DNA is suspended in liquid in the reaction zone.
  • the labeled and unlabeled dNTPs and other coupling reagents are flowed into the zone. Materials are removed from the zone through such a filter which retains the DNA chains.
  • the polymerase or other enzyme which is used to effect coupling is generally of a size to be retained by the membrane. This scheme works for chemical but not enzymatic deblocking, since in enzymatic deblocking the polymerase and phosphatase must be cycled separately through the cell.
  • the DNA can be immobilized on particles of resin or polymer microspheres and these particles retained within the chamber.
  • the filter material is unimportant as long as the DNA is attached to resin particles which are of a size that cannot penetrate the filter pores .
  • oligonucleotides or polynucleotides are linked through their 5' end to cellulose (Gilha , 1968; Clerici et al . 1979), Sephacryl (Langdale and Malcolm, 1985), or latex microspheres (Kremsky et al., 1987).
  • the DNA is available for interactions with other nucleic acids or proteins .
  • the DNA is coupled covalently to streptavidin-agarose beads by an alkylbiotinylated oligonucleotide (Kremsky et al., 1987).
  • the single-stranded DNA is coupled to DBM paper such as a filter in the presence of a protecting strand. After coupling, the protecting strand is released, leaving the immobilized template and priming site free for successive enzymatic reactions (Hansen et al . , 1987) .
  • This method and the other single-point methods described above are useful for immobilizing DNA while leaving it free for interactions with enzymes used in DNA sequencing- Examples
  • the organic layer is separated and the aqueous layer washed with 2 x 200 ml CH 2 C1 2
  • the combined CH-Cl- extracts are dried over magnesium sulfate (MgSO.), filtered and evaporated to dryness under vacuum at room temperature.
  • the crude 5 '-dimethoxytrityl-3 'thymidine H-phosphonate II is then treated with 2% benzenesulfonic acid in CH ⁇ Cl- -.methanol (MeOH) (7:3) (200 ml) for one hour.
  • the solution is washed with 10% sodium bicarbonate (NaHCO-) and water, dried over magnesium sulfate and evaporated to dryness.
  • the crude 3 '-thymidine- H-phosphonate III is recrystallized from ethanol/ether.
  • the mixtu-re is stirred for 12 hours at 4 C, neutralized with NaHCO.. solution,- and added to 150 ml water.
  • the aqueous solution is washed with benzene (2 x 100 ml) and ether ( 2 x 100 ml), and diluted to 0.8 liters with water and charged on a 2.5 x 50 cm column of DEAE-cellulose.
  • the products are eluted using a linear gradient of pH 8.5 ammonium bicarbonate solution (0.05 to 0.25 M) .
  • the fractions collected are analyzed by HPLC to determine the desired product-containing fractions, and these are evaporated to dryness under vacuum. The residue is repeatedly re-evaporated with water to remove salts .
  • the 5 '-monophosphate IV (16 mmole) is then dissolved in 30 ml of dimethylformamide (DMF) and treated with N,N'-carbonyldiimidazole (30 mmole) at room temperature for one hour.
  • the reaction is quenched by addition of 5 ml methanol, and 60 ml of a 0.5M solution of bis (tri-n-butyl-ammonium) pyrophosphate in DMF is added dropwise over 10 minutes.
  • the solution is diluted with water to 1 liter and treated with 100 ml of a solution of 0.1 M iodine (I-.) in 5% pyridine/water. After one hour, the solution is deposited on a DEAE-cellulose column from Sigma (5x50cm) or Sephradex from Pharmacia. The column is washed with water and eluted with triethylammonium bicarbonate solution
  • the 5 '-triphosphate-3 '-phosphate thymidine product V is obtained by evaporation of the appropriate fractions collected.
  • Example 3 Quartz Surface Immobilization of Subject DNA Four 25 microliter volume quartz cuvette reaction chambers are prepared. These chambers are configured like chamber 32 in Figure 3 with the exception that they use their inner walls as the surface to which the DNA is affixed. The inner surfaces are cleaned and dried.
  • Triethoxysilylpropylamine (5 microliter in 20 microliter CHC1-.) is added and held at 5°C for 120 minutes under anhydrous conditions . This couples the triethoxysilylpropylamine to the surface and gives an amine character to the surface.
  • the subject DNA is then attached to the amine surface.
  • This is carried out by first attaching a long chain alkyl amine (n-octylamine) to the base at the 5' end of the subject DNA molecule or to the base at the 5 ' end of a suitable primer, such as an M13 primer for example the 17-mer dGTAAAACGACGGCCAGT, and then joining the alkylamine to the aminopropyl ⁇ ilane surface groups by reaction with glutaraldehyde (1.5 equivalents, 25°C, 120 minutes).
  • a suitable primer such as an M13 primer for example the 17-mer dGTAAAACGACGGCCAGT
  • Other functional groups pendant to the base moiety or attached to the 5' position can also be used [for example: aldehydes or carboxylic acids (Kremsky et al) ] for covalent immobilization on derivatized quartz or glass surfaces.
  • Example 4 Incorporation of Labeled Nucleotide Analogs into DNA
  • the 25 microliter reaction zones are charged with a reaction mixture which contains three Units of Sequenase TM enzyme.
  • the reaction mixture also contains an appropriate buffer for this enzyme (20 mM Tris-HCl pH 7.5, 10 mM MgCl, 25 mM NaCl, 0.01 M dithiothreitol), the i single-stranded primed subject DNA is present at a concentration of approximately 0.1 M attached to the surface of the reaction chamber at its 5' end, (see Example 3), three unlabeled, 3 '-blocked deoxynucleotide triphosphate (dNTP) analogs at a concentration of 1.5 micromolar each, and one 3 '-blocked, fluorescently labeled dNTP analog of Example 2 at a concentration of 30 micromolar are each present in each of the four reaction zones. In each zone a different one of the four dNTPs is labeled.
  • the reaction
  • the identity of the added dNTP is determined by exciting the fluorophores present in the one cuvette which incorporated its fluorescently-labeled dNTP.
  • the fluorescent group is removed before measurement.
  • the 2 ,4-dinitrobenzenesulfenyl fluorescent blocking groups are removed with a deblocking reagent which consists of 0.1 M pyridine/pyridinium chloride buffer (pH 7.8) containing thiourea 0.05 M.
  • the deblocking reaction is allowed to proceed for one minute at 40 C-
  • the reaction chamber is then drained and washed twice with 100 mM Tris-HCl buffer, pH 6.5.
  • the release of the fluorescent blocking group is measured in the initial eluate from the reaction chamber using a flow-through cell. Depending on the cell in which the fluorescent group is present, the identity of the nucleotide which has been added to the DNA chain is determined.
  • the blocking group were a dansylcadaverine type ester such as in reaction scheme 4 , it could be removed by treatment with 50% methanol/50% water pH 10.0 for one minute.
  • Example 6 Enzymatic Deblocking
  • the blocking group can also be removed enzymaticall .
  • the deblocker fed into the reaction chamber contains 100 mM Tris-HCl (pH 6.5) 10 mM MgCl-, 5 mM 2-mercaptoethanol, and one Unit T4 polynucleotide kinase.
  • the reaction proceeds for one minute at a temperature of 37 C.
  • the 3 ' -phosphatase activity of T4 polynucleotide kinase converts 3 '-phosphate termini to 3 '-hydroxyl termini which then serve as primers for further synthesis.
  • a simple clock mechanism or microprocessor driven timer circuit can be used to actuate a plurality of electrically controlled valves in. sequence to add the various reagents for adding building blocks, deblocking and the like with the result that the sequence of the target DNA single strand can be obtained with minimum involvement of lab personnel .

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Molecular Biology (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Analytical Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Biotechnology (AREA)
  • Genetics & Genomics (AREA)
  • Biophysics (AREA)
  • Microbiology (AREA)
  • Pathology (AREA)
  • General Physics & Mathematics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

Instrument et procédé permettant de déterminer la séquence de nucléotides se trouvant dans une molécule d'ADN, sans l'emploi d'une étape d'électrophorèse sur gel. Le procédé emploie une séquence d'ADN monocaténaire amorçée inconnue, laquelle est immobilisée ou piégée à l'intérieur d'une chambre à l'aide d'une polymérase, de sorte que l'ADN complémentaire formé de manière séquentielle peut être contrôlé à chaque addition d'un nucléotide bloqué, par mesure de la présence d'un marqueur inoffensif sur des désoxyribonucléotides spécifiés. L'invention concerne également un procédé de détermination de la séquence de nucléotides d'ADN inconnue à l'aide de désoxynucléotides bloqués. Le dNTP (désoxyribonucléotide-triphosphate) bloqué comporte un marqueur inoffensif, de sorte que son identité peut être facilement déterminée. L'instrument et les procédés de l'invention permettent d'obtenir une détermination précise et rapide d'une séquence de nucléotides d'ADN sans électrophorèse sur gel.
EP19910900474 1989-10-26 1990-10-26 Sequen age d'adn Withdrawn EP0450060A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US42732189A 1989-10-26 1989-10-26
US427321 1999-10-26

Publications (1)

Publication Number Publication Date
EP0450060A1 true EP0450060A1 (fr) 1991-10-09

Family

ID=23694359

Family Applications (1)

Application Number Title Priority Date Filing Date
EP19910900474 Withdrawn EP0450060A1 (fr) 1989-10-26 1990-10-26 Sequen age d'adn

Country Status (3)

Country Link
EP (1) EP0450060A1 (fr)
CA (1) CA2044616A1 (fr)
WO (1) WO1991006678A1 (fr)

Families Citing this family (737)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5547839A (en) * 1989-06-07 1996-08-20 Affymax Technologies N.V. Sequencing of surface immobilized polymers utilizing microflourescence detection
WO1992016657A1 (fr) * 1991-03-13 1992-10-01 E.I. Du Pont De Nemours And Company Procede d'identification d'un nucleotide present dans un acide nucleique en une position definie
EP0514927A1 (fr) * 1991-05-24 1992-11-25 Walter Gilbert Procédé et dispositif de séquençage rapide des acides nucléiques
US5516633A (en) * 1991-08-15 1996-05-14 Amersham Life Science, Inc. DNA sequencing with a T7-type gene 6 exonuclease
GB9208733D0 (en) * 1992-04-22 1992-06-10 Medical Res Council Dna sequencing method
GB9210168D0 (en) * 1992-05-12 1992-06-24 Cemu Bioteknik Ab Method of sequencing dna
US5795714A (en) 1992-11-06 1998-08-18 Trustees Of Boston University Method for replicating an array of nucleic acid probes
US6194144B1 (en) 1993-01-07 2001-02-27 Sequenom, Inc. DNA sequencing by mass spectrometry
ATE220114T1 (de) * 1993-03-19 2002-07-15 Sequenom Inc Dns-sequenzbestimmung durch massenspektrometrie auf dem weg des abbaus mit exonuklease
FR2703052B1 (fr) * 1993-03-26 1995-06-02 Pasteur Institut Nouvelle méthode de séquençage d'acides nucléiques.
US6153379A (en) * 1993-06-22 2000-11-28 Baylor College Of Medicine Parallel primer extension approach to nucleic acid sequence analysis
US7001722B1 (en) 1993-06-22 2006-02-21 Baylor College Of Medicine Parallel primer extension approach to nucleic acid sequence analysis
EP1408122A3 (fr) * 1993-09-27 2004-10-06 Arch Development Corporation Procédés et compositions pour le séquençage efficace d'acide nucléique
US6401267B1 (en) 1993-09-27 2002-06-11 Radoje Drmanac Methods and compositions for efficient nucleic acid sequencing
GB9401200D0 (en) * 1994-01-21 1994-03-16 Medical Res Council Sequencing of nucleic acids
FR2718753B1 (fr) * 1994-04-15 1996-07-19 Pasteur Institut Procédé de comptage de mono-, di- et trinucléotides répétés dans un génome eucaryote et trousse permettant la mise en Óoeuvre de ce procédé.
US5604097A (en) 1994-10-13 1997-02-18 Spectragen, Inc. Methods for sorting polynucleotides using oligonucleotide tags
USRE43097E1 (en) 1994-10-13 2012-01-10 Illumina, Inc. Massively parallel signature sequencing by ligation of encoded adaptors
WO1998053300A2 (fr) 1997-05-23 1998-11-26 Lynx Therapeutics, Inc. Systeme et appareil destines au traitement sequentiel des analytes
SE9500342D0 (sv) * 1995-01-31 1995-01-31 Marek Kwiatkowski Novel chain terminators, the use thereof for nucleic acid sequencing and synthesis and a method of their preparation
AU5171696A (en) * 1995-02-27 1996-09-18 Ely Michael Rabani Device, compounds, algorithms, and methods of molecular characterization and manipulation with molecular parallelism
US5830655A (en) 1995-05-22 1998-11-03 Sri International Oligonucleotide sizing using cleavable primers
EP0745686A1 (fr) 1995-06-01 1996-12-04 Roche Diagnostics GmbH L'utilisation de l'activité de supression 3'-intrinsèque de l'ADN polymérase
EP0745688B1 (fr) * 1995-06-01 2007-02-14 Roche Diagnostics GmbH L'utilisation de l'activité de suppression 3'-intrinsique de l'ADN polymérase
GB9620209D0 (en) 1996-09-27 1996-11-13 Cemu Bioteknik Ab Method of sequencing DNA
US6133436A (en) * 1996-11-06 2000-10-17 Sequenom, Inc. Beads bound to a solid support and to nucleic acids
CA2270132A1 (fr) 1996-11-06 1998-05-14 Sequenom, Inc. Diagnostics de l'adn fondes sur la spectrometrie de masse
GB9626815D0 (en) 1996-12-23 1997-02-12 Cemu Bioteknik Ab Method of sequencing DNA
US7622294B2 (en) 1997-03-14 2009-11-24 Trustees Of Tufts College Methods for detecting target analytes and enzymatic reactions
US20030027126A1 (en) 1997-03-14 2003-02-06 Walt David R. Methods for detecting target analytes and enzymatic reactions
EP1498494A3 (fr) 1997-04-01 2007-06-20 Solexa Ltd. Procédé de séquencage d'acides nucléiques
EP2327797B1 (fr) 1997-04-01 2015-11-25 Illumina Cambridge Limited Methode de séquençage d'acide nucléique
ATE225858T1 (de) * 1997-07-28 2002-10-15 Medical Biosystems Ltd Sequenzanalyse von nukleinsäuren
US7875440B2 (en) * 1998-05-01 2011-01-25 Arizona Board Of Regents Method of determining the nucleotide sequence of oligonucleotides and DNA molecules
WO1999057321A1 (fr) * 1998-05-01 1999-11-11 Arizona Board Of Regents Procede de determination de la sequence nucleotidique d'oligonucleotides et de molecules d'adn
US6780591B2 (en) * 1998-05-01 2004-08-24 Arizona Board Of Regents Method of determining the nucleotide sequence of oligonucleotides and DNA molecules
AR021833A1 (es) 1998-09-30 2002-08-07 Applied Research Systems Metodos de amplificacion y secuenciacion de acido nucleico
AU1818600A (en) 1998-11-16 2000-06-05 Genway Biotech, Inc. Generation of antibodies using polynucleotide vaccination in avian species
WO2000036151A1 (fr) 1998-12-14 2000-06-22 Li-Cor, Inc. Essai heterogene pour detection des pyrophosphates
NO986133D0 (no) 1998-12-23 1998-12-23 Preben Lexow FremgangsmÕte for DNA-sekvensering
DK1159453T3 (da) * 1999-03-10 2008-10-06 Asm Scient Inc Fremgangsmåde til direkte nukleinsyresekventering
WO2000058507A1 (fr) * 1999-03-30 2000-10-05 Solexa Ltd. Sequençage de polynucleotide
US20060275782A1 (en) 1999-04-20 2006-12-07 Illumina, Inc. Detection of nucleic acid reactions on bead arrays
CA2374390A1 (fr) 1999-05-20 2000-12-14 Illumina, Inc. Decodage combinatoire de jeux d'acides nucleiques aleatoires
US7501245B2 (en) 1999-06-28 2009-03-10 Helicos Biosciences Corp. Methods and apparatuses for analyzing polynucleotide sequences
US6818395B1 (en) 1999-06-28 2004-11-16 California Institute Of Technology Methods and apparatus for analyzing polynucleotide sequences
US6908736B1 (en) 1999-10-06 2005-06-21 Medical Biosystems, Ltd. DNA sequencing method
GB9923644D0 (en) 1999-10-06 1999-12-08 Medical Biosystems Ltd DNA sequencing
AU3015801A (en) * 1999-12-23 2001-07-09 Axaron Bioscience Ag Method for carrying out the parallel sequencing of a nucleic acid mixture on a surface
US7582420B2 (en) 2001-07-12 2009-09-01 Illumina, Inc. Multiplex nucleic acid reactions
US7611869B2 (en) 2000-02-07 2009-11-03 Illumina, Inc. Multiplexed methylation detection methods
JP3442338B2 (ja) 2000-03-17 2003-09-02 株式会社日立製作所 Ｄｎａ分析装置、ｄｎａ塩基配列決定装置、ｄｎａ塩基配列決定方法、および反応モジュール
EP1182267B1 (fr) * 2000-03-30 2012-01-18 Toyota Jidosha Kabushiki Kaisha Procede pour determiner une sequence de base d'une molecule individuelle d'acide nucleique
AU2001254771A1 (en) * 2000-04-03 2001-10-15 Axaron Bioscience Ag Novel method for the parallel sequencing of a nucleic acid mixture on a surface
GB0016472D0 (en) * 2000-07-05 2000-08-23 Amersham Pharm Biotech Uk Ltd Sequencing method and apparatus
US6664079B2 (en) 2000-10-06 2003-12-16 The Trustees Of Columbia University In The City Of New York Massive parallel method for decoding DNA and RNA
US9708358B2 (en) 2000-10-06 2017-07-18 The Trustees Of Columbia University In The City Of New York Massive parallel method for decoding DNA and RNA
EP1354064A2 (fr) 2000-12-01 2003-10-22 Visigen Biotechnologies, Inc. Synthese d'acides nucleiques d'enzymes, et compositions et methodes modifiant la fidelite d'incorporation de monomeres
AR031640A1 (es) 2000-12-08 2003-09-24 Applied Research Systems Amplificacion isotermica de acidos nucleicos en un soporte solido
EP1368497A4 (fr) 2001-03-12 2007-08-15 California Inst Of Techn Procedes et appareil d'analyse de sequences de polynucleotide par extension de base asynchrone
US6653082B2 (en) 2001-05-17 2003-11-25 Baylor College Of Medicine Substrate-bound cleavage assay for nucleic acid analysis
US7057026B2 (en) * 2001-12-04 2006-06-06 Solexa Limited Labelled nucleotides
GB0129012D0 (en) 2001-12-04 2002-01-23 Solexa Ltd Labelled nucleotides
WO2003102216A2 (fr) 2002-05-31 2003-12-11 Diversa Corporation Systemes multiplexes conçus pour le sequençage d'acide nucleique
JP4106977B2 (ja) 2002-06-21 2008-06-25 株式会社日立製作所 分析チップ及び分析装置
US7074597B2 (en) 2002-07-12 2006-07-11 The Trustees Of Columbia University In The City Of New York Multiplex genotyping using solid phase capturable dideoxynucleotides and mass spectrometry
US11008359B2 (en) 2002-08-23 2021-05-18 Illumina Cambridge Limited Labelled nucleotides
US7414116B2 (en) 2002-08-23 2008-08-19 Illumina Cambridge Limited Labelled nucleotides
ES2864086T3 (es) 2002-08-23 2021-10-13 Illumina Cambridge Ltd Nucleótidos etiquetados
AU2003259350A1 (en) 2002-08-23 2004-03-11 Solexa Limited Modified nucleotides for polynucleotide sequencing
EP1725572B1 (fr) 2003-11-05 2017-05-31 AGCT GmbH Composes nucleotidiques macromoleculaires et leurs procedes d'utilisation
US7169560B2 (en) 2003-11-12 2007-01-30 Helicos Biosciences Corporation Short cycle methods for sequencing polynucleotides
EP1716254B1 (fr) 2004-02-19 2010-04-07 Helicos Biosciences Corporation Procedes pour analyser des sequences de polynucleotides
CA2557818A1 (fr) 2004-03-03 2005-09-15 The Trustees Of Columbia University In The City Of New York Nucleotides fluorescents photoclivables destines a sequencer de l'adn sur une puce construite par chimie de couplage specifique au site
JP2008512084A (ja) 2004-05-25 2008-04-24 ヘリコス バイオサイエンシーズ コーポレイション 核酸の配列決定のための方法およびデバイス
US7476734B2 (en) 2005-12-06 2009-01-13 Helicos Biosciences Corporation Nucleotide analogs
EP1790202A4 (fr) * 2004-09-17 2013-02-20 Pacific Biosciences California Appareil et procede d'analyse de molecules
US7220549B2 (en) 2004-12-30 2007-05-22 Helicos Biosciences Corporation Stabilizing a nucleic acid for nucleic acid sequencing
US7482120B2 (en) 2005-01-28 2009-01-27 Helicos Biosciences Corporation Methods and compositions for improving fidelity in a nucleic acid synthesis reaction
CA2593855A1 (fr) * 2005-01-31 2006-08-10 Pacific Biosciences Of California, Inc. Utilisation de terminateur d'extension reversible dans le sequencage d'acides nucleiques
EP2241637A1 (fr) 2005-02-01 2010-10-20 AB Advanced Genetic Analysis Corporation Sequençage nucléotidique comprenant des cycles successifs de formation duplex
EP2230315A1 (fr) 2005-02-01 2010-09-22 AB Advanced Genetic Analysis Corporation Sequencage nucléotidique comprenant des cycles sucessifs de fromation duplex
US9169510B2 (en) 2005-06-21 2015-10-27 The Trustees Of Columbia University In The City Of New York Pyrosequencing methods and related compositions
US7805081B2 (en) 2005-08-11 2010-09-28 Pacific Biosciences Of California, Inc. Methods and systems for monitoring multiple optical signals from a single source
GB0517097D0 (en) 2005-08-19 2005-09-28 Solexa Ltd Modified nucleosides and nucleotides and uses thereof
US7666593B2 (en) 2005-08-26 2010-02-23 Helicos Biosciences Corporation Single molecule sequencing of captured nucleic acids
US7405281B2 (en) 2005-09-29 2008-07-29 Pacific Biosciences Of California, Inc. Fluorescent nucleotide analogs and uses therefor
US7763423B2 (en) 2005-09-30 2010-07-27 Pacific Biosciences Of California, Inc. Substrates having low density reactive groups for monitoring enzyme activity
WO2007053719A2 (fr) 2005-10-31 2007-05-10 The Trustees Of Columbia University In The City Of New York Analogues nucleotidiques fluorescents de 3'-o-allyl-dntp-allyl-fluorophore chimiquement clivables et procedes associes
GB2446084B (en) 2005-10-31 2011-03-02 Univ Columbia Synthesis of four color 3-o-allyl modified photocleavable fluorescent nucleotides and related methods
GB0522310D0 (en) 2005-11-01 2005-12-07 Solexa Ltd Methods of preparing libraries of template polynucleotides
GB0524069D0 (en) 2005-11-25 2006-01-04 Solexa Ltd Preparation of templates for solid phase amplification
US7998717B2 (en) 2005-12-02 2011-08-16 Pacific Biosciences Of California, Inc. Mitigation of photodamage in analytical reactions
US7715001B2 (en) 2006-02-13 2010-05-11 Pacific Biosciences Of California, Inc. Methods and systems for simultaneous real-time monitoring of optical signals from multiple sources
US7995202B2 (en) 2006-02-13 2011-08-09 Pacific Biosciences Of California, Inc. Methods and systems for simultaneous real-time monitoring of optical signals from multiple sources
US7692783B2 (en) 2006-02-13 2010-04-06 Pacific Biosciences Of California Methods and systems for simultaneous real-time monitoring of optical signals from multiple sources
US7397546B2 (en) 2006-03-08 2008-07-08 Helicos Biosciences Corporation Systems and methods for reducing detected intensity non-uniformity in a laser beam
US8975216B2 (en) 2006-03-30 2015-03-10 Pacific Biosciences Of California Articles having localized molecules disposed thereon and methods of producing same
US7563574B2 (en) 2006-03-31 2009-07-21 Pacific Biosciences Of California, Inc. Methods, systems and compositions for monitoring enzyme activity and applications thereof
WO2007135368A2 (fr) 2006-05-18 2007-11-29 Solexa Limited Composés de type teinte et application de leurs conjugués marqués
CN101495656B (zh) 2006-06-07 2017-02-08 纽约哥伦比亚大学理事会 采用带修饰的核苷酸通过纳米通道进行dna序列测定
AU2007289057C1 (en) 2006-09-01 2014-01-16 Pacific Biosciences Of California, Inc. Substrates, systems and methods for analyzing materials
US8207509B2 (en) 2006-09-01 2012-06-26 Pacific Biosciences Of California, Inc. Substrates, systems and methods for analyzing materials
US8399188B2 (en) 2006-09-28 2013-03-19 Illumina, Inc. Compositions and methods for nucleotide sequencing
US7883869B2 (en) 2006-12-01 2011-02-08 The Trustees Of Columbia University In The City Of New York Four-color DNA sequencing by synthesis using cleavable fluorescent nucleotide reversible terminators
EP2121983A2 (fr) 2007-02-02 2009-11-25 Illumina Cambridge Limited Procedes pour indexer des echantillons et sequencer de multiples matrices nucleotidiques
US11940413B2 (en) 2007-02-05 2024-03-26 IsoPlexis Corporation Methods and devices for sequencing nucleic acids in smaller batches
US8551704B2 (en) 2007-02-16 2013-10-08 Pacific Biosciences Of California, Inc. Controllable strand scission of mini circle DNA
EP2183388A4 (fr) 2007-07-26 2010-09-08 Pacific Biosciences California Séquençage moléculaire redondant
WO2009029728A1 (fr) 2007-08-29 2009-03-05 Applied Biosystems Inc. Procédé de séquençage alternatif d'acide nucléique
CA2700838A1 (fr) 2007-09-28 2009-04-09 Pacific Biosciences Of California, Inc. Amplification sans erreur d'adn pour le sequencage de clones
US7960116B2 (en) 2007-09-28 2011-06-14 Pacific Biosciences Of California, Inc. Nucleic acid sequencing methods and systems
WO2009051807A1 (fr) 2007-10-19 2009-04-23 The Trustees Of Columbia University In The City Of New York Conception et synthèse de nucléotides fluorescents clivables en tant que terminateurs réversibles pour le séquençage de l'adn par synthèse
EP3431615A3 (fr) 2007-10-19 2019-02-20 The Trustees of Columbia University in the City of New York Séquençage d'adn avec des terminateurs nucléotidiques réversibles non fluorescents et terminateurs nucléotidiques modifiées à étiquette clivable
US8017338B2 (en) 2007-11-20 2011-09-13 Life Technologies Corporation Reversible di-nucleotide terminator sequencing
AU2009204461A1 (en) 2008-01-10 2009-07-16 Pacific Biosciences Of California, Inc. Methods and systems for analysis of fluorescent reactions with modulated excitation
WO2009097368A2 (fr) 2008-01-28 2009-08-06 Complete Genomics, Inc. Procédés et compositions pour un appel de base efficace dans des réactions de séquençage
WO2009102470A2 (fr) 2008-02-12 2009-08-20 Pacific Biosciences Of California, Inc. Compositions et procédés utilisables dans des réactions analytiques
US9017973B2 (en) 2008-03-19 2015-04-28 Intelligent Biosystems, Inc. Methods and compositions for incorporating nucleotides
US8143030B2 (en) 2008-09-24 2012-03-27 Pacific Biosciences Of California, Inc. Intermittent detection during analytical reactions
EP3045542B1 (fr) 2008-03-28 2016-11-16 Pacific Biosciences of California, Inc. Procédés pour le séquençage d'acide nucléique
WO2009120374A2 (fr) 2008-03-28 2009-10-01 Pacific Biosciences Of California, Inc. Procédés et compositions pour la préparation d’échantillon d’acide nucléique
US8628940B2 (en) 2008-09-24 2014-01-14 Pacific Biosciences Of California, Inc. Intermittent detection during analytical reactions
EP2294076B1 (fr) 2008-05-27 2017-03-08 TriLink BioTechnologies Nucléosides 5 -triphosphates modifiés chimiquement pour la réplication initiée thermiquement d'un acide nucléique
US8198023B2 (en) 2008-08-05 2012-06-12 Pacific Biosciences Of California, Inc. Prevention and alleviation of steric hindrance during single molecule nucleic acid synthesis by a polymerase
WO2010027497A2 (fr) 2008-09-05 2010-03-11 Pacific Biosciences Of California, Inc Préparations, compositions et procédés de séquençage d'acides nucléiques
US8383345B2 (en) 2008-09-12 2013-02-26 University Of Washington Sequence tag directed subassembly of short sequencing reads into long sequencing reads
WO2010033193A2 (fr) 2008-09-16 2010-03-25 Pacific Biosciences Of California, Inc. Substrats et systèmes optiques et leurs procédés d'utilisation
US8921046B2 (en) 2008-09-19 2014-12-30 Pacific Biosciences Of California, Inc. Nucleic acid sequence analysis
US8481264B2 (en) 2008-09-19 2013-07-09 Pacific Biosciences Of California, Inc. Immobilized nucleic acid complexes for sequence analysis
US8383369B2 (en) 2008-09-24 2013-02-26 Pacific Biosciences Of California, Inc. Intermittent detection during analytical reactions
US8728764B2 (en) 2008-10-02 2014-05-20 Illumina Cambridge Limited Nucleic acid sample enrichment for sequencing applications
US8541207B2 (en) 2008-10-22 2013-09-24 Illumina, Inc. Preservation of information related to genomic DNA methylation
US8486865B2 (en) 2008-11-03 2013-07-16 The Regents Of The University Of California Methods for detecting modification resistant nucleic acids
US8252910B2 (en) 2008-11-19 2012-08-28 Pacific Biosciences Of California, Inc. Modular nucleotide compositions and uses therefor
WO2010059235A2 (fr) 2008-11-20 2010-05-27 Pacific Biosciences Of California, Inc. Algorithmes de détermination de séquence
US8993230B2 (en) 2008-12-04 2015-03-31 Pacific Biosciences of Californ, Inc. Asynchronous sequencing of biological polymers
US9175338B2 (en) 2008-12-11 2015-11-03 Pacific Biosciences Of California, Inc. Methods for identifying nucleic acid modifications
US12527665B2 (en) 2008-12-11 2026-01-20 Pacific Biosciences Of California, Inc. Methods for identifying modified bases in nucleic acid templates
EP2370598B1 (fr) 2008-12-11 2017-02-15 Pacific Biosciences Of California, Inc. Classification de matrices d'acides nucléiques
EP2379748A4 (fr) 2008-12-23 2012-08-29 Illumina Inc Libération multibase pour lectures longues dans le séquençage par des protocoles de synthèse
US20100255487A1 (en) 2009-03-27 2010-10-07 Life Technologies Corporation Methods and apparatus for single molecule sequencing using energy transfer detection
EP2425023B1 (fr) 2009-04-27 2015-12-23 Pacific Biosciences of California, Inc. Procédés et systèmes de séquençage en temps réel
US8632975B2 (en) 2009-06-05 2014-01-21 Life Technologies Corporation Nucleotide transient binding for sequencing methods
US8501406B1 (en) 2009-07-14 2013-08-06 Pacific Biosciences Of California, Inc. Selectively functionalized arrays
US8182994B2 (en) 2009-09-15 2012-05-22 Illumina Cambridge Limited Centroid markers for image analysis of high denisty clusters in complex polynucleotide sequencing
WO2011050938A1 (fr) 2009-10-26 2011-05-05 Genovoxx Gmbh Conjugués de nucléotides et méthodes d'utilisation associées
PL2531880T3 (pl) 2010-02-01 2017-09-29 Illumina Inc. Sposoby ogniskowania oraz układy optyczne i zespoły optyczne realizujące te sposoby
US8518643B2 (en) 2010-02-04 2013-08-27 Pacific Biosciences Of California, Inc. Method to improve single molecule analyses
US9678055B2 (en) 2010-02-08 2017-06-13 Genia Technologies, Inc. Methods for forming a nanopore in a lipid bilayer
US9605307B2 (en) 2010-02-08 2017-03-28 Genia Technologies, Inc. Systems and methods for forming a nanopore in a lipid bilayer
US8324914B2 (en) 2010-02-08 2012-12-04 Genia Technologies, Inc. Systems and methods for characterizing a molecule
US8994946B2 (en) 2010-02-19 2015-03-31 Pacific Biosciences Of California, Inc. Integrated analytical system and method
US9410891B2 (en) 2010-02-19 2016-08-09 Pacific Biosciences Of California, Inc. Optics collection and detection system and method
CN202281746U (zh) 2010-03-06 2012-06-20 伊鲁米那股份有限公司 检测来自样品光信号的测定设备及其光学组件和光学系统
US10787701B2 (en) 2010-04-05 2020-09-29 Prognosys Biosciences, Inc. Spatially encoded biological assays
DK2556171T3 (en) 2010-04-05 2015-12-14 Prognosys Biosciences Inc Spatially CODED BIOLOGICAL ASSAYS
US20190300945A1 (en) 2010-04-05 2019-10-03 Prognosys Biosciences, Inc. Spatially Encoded Biological Assays
WO2011159942A1 (fr) 2010-06-18 2011-12-22 Illumina, Inc. Sondes conformationnelles et procédés de séquençage d'acides nucléiques
US8318094B1 (en) 2010-06-18 2012-11-27 Pacific Biosciences Of California, Inc. Substrate analysis systems
US8834847B2 (en) 2010-08-12 2014-09-16 Pacific Biosciences Of California, Inc. Photodamage mitigation compounds and systems
US8465922B2 (en) 2010-08-26 2013-06-18 Pacific Biosciences Of California, Inc. Methods and systems for monitoring reactions
US9029103B2 (en) 2010-08-27 2015-05-12 Illumina Cambridge Limited Methods for sequencing polynucleotides
US8483969B2 (en) 2010-09-17 2013-07-09 Illuminia, Inc. Variation analysis for multiple templates on a solid support
WO2012050920A1 (fr) 2010-09-29 2012-04-19 Illumina, Inc. Compositions et procédés de séquençage d'acides nucléiques
EP2633069B1 (fr) 2010-10-26 2015-07-01 Illumina, Inc. Procédés de séquençage
US8575071B2 (en) 2010-11-03 2013-11-05 Illumina, Inc. Reducing adapter dimer formation
US9074251B2 (en) 2011-02-10 2015-07-07 Illumina, Inc. Linking sequence reads using paired code tags
CA2821299C (fr) 2010-11-05 2019-02-12 Frank J. Steemers Liaison entre des lectures de sequences a l'aide de codes marqueurs apparies
EP2643484A4 (fr) 2010-11-22 2014-04-16 Univ California Procédés d'identification d'un transcrit cellulaire naissant d'arn
US9121059B2 (en) 2010-12-22 2015-09-01 Genia Technologies, Inc. Nanopore-based single molecule characterization
US8951781B2 (en) 2011-01-10 2015-02-10 Illumina, Inc. Systems, methods, and apparatuses to image a sample for biological or chemical analysis
US9581563B2 (en) 2011-01-24 2017-02-28 Genia Technologies, Inc. System for communicating information from an array of sensors
US9110478B2 (en) 2011-01-27 2015-08-18 Genia Technologies, Inc. Temperature regulation of measurement arrays
EP2670864B1 (fr) 2011-01-31 2017-03-08 Illumina, Inc. Procédés pour réduire les dommages occasionnés aux acides nucléiques
JP6017458B2 (ja) 2011-02-02 2016-11-02 ユニヴァーシティ・オブ・ワシントン・スルー・イッツ・センター・フォー・コマーシャリゼーション 大量並列連続性マッピング
EP2689028B1 (fr) 2011-03-23 2017-08-30 Pacific Biosciences Of California, Inc. Isolement de complexes polymérase-acide nucléique et charge sur des substrats
EP2694686B2 (fr) 2011-04-06 2023-07-19 The University of Chicago Composition et procédés se rapportant à la modification de la 5-méthylcytosine (5-mc)
GB201106254D0 (en) 2011-04-13 2011-05-25 Frisen Jonas Method and product
US9994894B2 (en) 2011-04-27 2018-06-12 Agct Gmbh Method and components for detecting nucleic acid chains
US20150086981A1 (en) 2011-05-04 2015-03-26 Genovoxx Gmbh Nucleoside-triphosphate conjugate and methods for the use thereof
US9624539B2 (en) 2011-05-23 2017-04-18 The Trustees Of Columbia University In The City Of New York DNA sequencing by synthesis using Raman and infrared spectroscopy detection
US8778848B2 (en) 2011-06-09 2014-07-15 Illumina, Inc. Patterned flow-cells useful for nucleic acid analysis
WO2013049135A1 (fr) 2011-09-26 2013-04-04 Gen-Probe Incorporated Algorithmes de détermination de séquences
US10378051B2 (en) 2011-09-29 2019-08-13 Illumina Cambridge Limited Continuous extension and deblocking in reactions for nucleic acids synthesis and sequencing
US9347900B2 (en) 2011-10-14 2016-05-24 Pacific Biosciences Of California, Inc. Real-time redox sequencing
EP3305400A3 (fr) 2011-10-28 2018-06-06 Illumina, Inc. Système et procédé de fabrication de microréseau
EP2776165A2 (fr) 2011-11-07 2014-09-17 Illumina, Inc. Appareils de séquençage intégré et procédés d'utilisation
EP2788499B1 (fr) 2011-12-09 2016-01-13 Illumina, Inc. Base de numération étendue pour étiquettes polymères
US9279154B2 (en) 2011-12-21 2016-03-08 Illumina, Inc. Apparatus and methods for kinetic analysis and determination of nucleic acid sequences
US9238836B2 (en) 2012-03-30 2016-01-19 Pacific Biosciences Of California, Inc. Methods and compositions for sequencing modified nucleic acids
WO2013117595A2 (fr) 2012-02-07 2013-08-15 Illumina Cambridge Limited Enrichissement et amplification ciblés d'acides nucléiques sur un support
US9062091B2 (en) 2012-02-15 2015-06-23 Pacific Biosciences Of California, Inc. Polymerase enzyme substrates with protein shield
US8986629B2 (en) 2012-02-27 2015-03-24 Genia Technologies, Inc. Sensor circuit for controlling, detecting, and measuring a molecular complex
NO2694769T3 (fr) 2012-03-06 2018-03-03
US20130261984A1 (en) 2012-03-30 2013-10-03 Illumina, Inc. Methods and systems for determining fetal chromosomal abnormalities
CN204832037U (zh) 2012-04-03 2015-12-02 伊鲁米那股份有限公司 检测设备
US20130274148A1 (en) 2012-04-11 2013-10-17 Illumina, Inc. Portable genetic detection and analysis system and method
US9175348B2 (en) 2012-04-24 2015-11-03 Pacific Biosciences Of California, Inc. Identification of 5-methyl-C in nucleic acid templates
US9012022B2 (en) 2012-06-08 2015-04-21 Illumina, Inc. Polymer coatings
US8895249B2 (en) 2012-06-15 2014-11-25 Illumina, Inc. Kinetic exclusion amplification of nucleic acid libraries
WO2013188841A1 (fr) 2012-06-15 2013-12-19 Genia Technologies, Inc. Configuration de puce et séquençage d'acide nucléique à haute précision
US9372308B1 (en) 2012-06-17 2016-06-21 Pacific Biosciences Of California, Inc. Arrays of integrated analytical devices and methods for production
EP2870264A4 (fr) 2012-07-03 2016-03-02 Sloan Kettering Inst Cancer Évaluation quantitative de la reconstitution du répertoire des cellules t chez l'homme après une greffe allogénique de cellules souches hématopoïétiques
US9695416B2 (en) 2012-07-18 2017-07-04 Siemens Healthcare Diagnostics Inc. Method of normalizing biological samples
NL2017959B1 (en) 2016-12-08 2018-06-19 Illumina Inc Cartridge assembly
CA3040684C (fr) 2012-08-20 2023-02-07 Hod Finkelstein Procede et systeme de sequencage reposant sur la duree de vie de fluorescence
WO2014060483A1 (fr) 2012-10-17 2014-04-24 Spatial Transcriptomics Ab Procédés et produit d'optimisation de la détection localisée ou spatiale de l'expression génique dans un échantillon de tissu
US9181583B2 (en) 2012-10-23 2015-11-10 Illumina, Inc. HLA typing using selective amplification and sequencing
US10206911B2 (en) 2012-10-26 2019-02-19 Memorial Sloan-Kettering Cancer Center Androgen receptor variants and methods for making and using
US9605309B2 (en) 2012-11-09 2017-03-28 Genia Technologies, Inc. Nucleic acid sequencing using tags
WO2014099776A1 (fr) 2012-12-18 2014-06-26 Pacific Biosciences Of California, Inc. Éclairage de dispositifs analytiques optiques
US9683230B2 (en) 2013-01-09 2017-06-20 Illumina Cambridge Limited Sample preparation on a solid support
US9805407B2 (en) 2013-01-25 2017-10-31 Illumina, Inc. Methods and systems for using a cloud computing environment to configure and sell a biological sample preparation cartridge and share related data
US9759711B2 (en) 2013-02-05 2017-09-12 Genia Technologies, Inc. Nanopore arrays
US9624540B2 (en) 2013-02-22 2017-04-18 Pacific Biosciences Of California, Inc. Integrated illumination of optical analytical devices
US9512422B2 (en) 2013-02-26 2016-12-06 Illumina, Inc. Gel patterned surfaces
EP2964785B8 (fr) 2013-03-04 2020-10-21 Fry Laboratories LLC Procédé et trousse pour la caractérisation de micro-organismes
WO2014142841A1 (fr) 2013-03-13 2014-09-18 Illumina, Inc. Dispositifs fluidiques multicouches et leurs procédés de fabrication
DK3553175T3 (da) 2013-03-13 2021-08-23 Illumina Inc Fremgangsmåde til fremstilling af et nukleinsyresekvenseringsbibliotek
WO2014142921A1 (fr) 2013-03-14 2014-09-18 Illumina, Inc. Polymérases modifiées pour l'incorporation améliorée d'analogues nucléotidiques
US10648026B2 (en) 2013-03-15 2020-05-12 The Trustees Of Columbia University In The City Of New York Raman cluster tagged molecules for biological imaging
US20140274747A1 (en) 2013-03-15 2014-09-18 Illumina, Inc. Super resolution imaging
US9193998B2 (en) 2013-03-15 2015-11-24 Illumina, Inc. Super resolution imaging
CN105377869B (zh) 2013-03-15 2018-07-10 伊鲁米纳剑桥有限公司 修饰的核苷或核苷酸
EP2971071B1 (fr) 2013-03-15 2018-02-28 Illumina, Inc. Nucléotides à liaison enzymatique
EP3008211A4 (fr) 2013-06-11 2016-12-21 Courtagen Life Sciences Inc Méthodes et nécessaires permettant de traiter et de classer des individus risquant de souffrir ou souffrant d'un changement de fonction de trap1
US9868979B2 (en) 2013-06-25 2018-01-16 Prognosys Biosciences, Inc. Spatially encoded biological assays using a microfluidic device
MX363806B (es) 2013-07-01 2019-04-03 Illumina Inc Funcionalización de superficies sin catalizador e injerto de polímeros..
EP3241913B1 (fr) 2013-07-03 2019-02-20 Illumina, Inc. Système de séquençage par synthèse orthogonale
WO2015021079A1 (fr) 2013-08-05 2015-02-12 Pacific Biosciences Of California, Inc. Composés réactifs fluorescents protégés
KR102043320B1 (ko) 2013-08-08 2019-11-11 일루미나, 인코포레이티드 플로우 셀로의 시약 전달을 위한 유체 시스템
EP3038649B1 (fr) 2013-08-26 2019-09-25 The Translational Genomics Research Institute Analyse de lecture à chevauchement monomoléculaire pour la détection de mutation variante mineure dans des échantillons de pathogènes
US9551697B2 (en) 2013-10-17 2017-01-24 Genia Technologies, Inc. Non-faradaic, capacitively coupled measurement in a nanopore cell array
US9567630B2 (en) 2013-10-23 2017-02-14 Genia Technologies, Inc. Methods for forming lipid bilayers on biochips
JP6461943B2 (ja) 2013-10-23 2019-01-30 ジェニア・テクノロジーズ・インコーポレイテッド ナノポアを備えた高速分子検知
US9416414B2 (en) 2013-10-24 2016-08-16 Pacific Biosciences Of California, Inc. Delaying real-time sequencing
US10540783B2 (en) 2013-11-01 2020-01-21 Illumina, Inc. Image analysis useful for patterned objects
US9983135B2 (en) 2013-11-17 2018-05-29 Quantum-Si Incorporated Active-source-pixel, integrated device for rapid analysis of biological and chemical specimens
EP2876166B1 (fr) 2013-11-20 2016-12-14 Roche Diagnostics GmbH Nouveaux composés pour le séquençage par synthèse
HRP20201397T1 (hr) 2013-12-03 2020-11-27 Illumina, Inc. Postupak i sustavi za analizu slikovnih podataka
NZ720871A (en) 2013-12-10 2020-03-27 Illumina Inc Biosensors for biological or chemical analysis and methods of manufacturing the same
CN106414765A (zh) 2013-12-20 2017-02-15 Illumina公司 在片段化的基因组dna样品中保留基因组连接信息
KR102475314B1 (ko) 2013-12-23 2022-12-06 일루미나, 인코포레이티드 광 방출의 검출을 개선시키기 위한 구조화 기판 및 이와 관련한 방법
WO2015103225A1 (fr) 2013-12-31 2015-07-09 Illumina, Inc. Cuve à circulation adressable utilisant des électrodes à motif
US9677132B2 (en) 2014-01-16 2017-06-13 Illumina, Inc. Polynucleotide modification on solid support
US9926598B2 (en) 2014-01-16 2018-03-27 Illumina, Inc. Amplicon preparation and sequencing on solid supports
ES2866044T3 (es) 2014-02-18 2021-10-19 Illumina Inc Métodos y composiciones para la elaboración de perfiles de ADN
CN110982689A (zh) 2014-03-11 2020-04-10 伊鲁米那股份有限公司 一次性的集成微流体盒及其制备和使用的方法
FR3020071B1 (fr) 2014-04-17 2017-12-22 Dna Script Procede de synthese d'acides nucleiques, notamment d'acides nucleiques de grande longueur, utilisation du procede et kit pour la mise en œuvre du procede
CN106459967A (zh) 2014-04-29 2017-02-22 Illumina公司 使用模板转换和标签化的多重单细胞基因表达分析
EP3143161B1 (fr) 2014-05-16 2021-04-21 Illumina, Inc. Techniques de synthèse d'acide nucléique
AU2015267189B2 (en) 2014-05-27 2019-12-05 Illumina, Inc. Systems and methods for biochemical analysis including a base instrument and a removable cartridge
CA2950298C (fr) 2014-06-03 2024-01-09 Illumina, Inc. Compositions, systemes et procedes pour detecter des evenements a l'aide d'attaches ancrees ou adjacentes a des nanopores
US20150353989A1 (en) 2014-06-09 2015-12-10 Illumina Cambridge Limited Sample preparation for nucleic acid amplification
CN106715693A (zh) 2014-06-13 2017-05-24 伊卢米纳剑桥有限公司 用于制备序列文库的方法和组合物
US10829814B2 (en) 2014-06-19 2020-11-10 Illumina, Inc. Methods and compositions for single cell genomics
US10017759B2 (en) 2014-06-26 2018-07-10 Illumina, Inc. Library preparation of tagged nucleic acid
ES2788949T3 (es) 2014-06-27 2020-10-23 Illumina Inc Polimerasas modificadas para la incorporación mejorada de análogos de nucleótidos
ES2713153T3 (es) 2014-06-30 2019-05-20 Illumina Inc Métodos y composiciones que utilizan la transposición unilateral
AU2015289828B2 (en) 2014-07-15 2019-12-19 Illumina, Inc. Biochemically activated electronic device
CN112941065A (zh) 2014-07-21 2021-06-11 亿明达股份有限公司 使用crispr-cas系统的多核苷酸富集
JP6812341B2 (ja) 2014-08-08 2021-01-13 クアンタム−エスアイ インコーポレイテッドＱｕａｎｔｕｍ−Ｓｉ Ｉｎｃｏｒｐｏｒａｔｅｄ 分子の探索、検出及び解析のための光学システム及びアッセイチップ
CA2957546A1 (fr) 2014-08-08 2016-02-11 Quantum-Si Incorporated Dispositif integre, avec source lumineuse externe, pour sonder, detecter, et analyser des molecules
GB201414098D0 (en) 2014-08-08 2014-09-24 Illumina Cambridge Ltd Modified nucleotide linkers
BR112017002485B1 (pt) 2014-08-08 2020-12-01 Quantum-Si Incorporated circuito integrado e método de fotodetecção
US10174363B2 (en) 2015-05-20 2019-01-08 Quantum-Si Incorporated Methods for nucleic acid sequencing
WO2016026924A1 (fr) 2014-08-21 2016-02-25 Illumina Cambridge Limited Fonctionnalisation de surface réversible
TWI692633B (zh) 2014-08-27 2020-05-01 美商加州太平洋生物科學公司 整合式分析裝置之陣列
FR3025201B1 (fr) 2014-09-02 2018-10-12 Dna Script Nucleotides modifies pour la synthese d'acides nucleiques, un kit renfermant de tels nucleotides et leur utilisation pour la production de genes ou sequences d'acides nucleiques synthetiques
WO2016040602A1 (fr) 2014-09-11 2016-03-17 Epicentre Technologies Corporation Séquence au bisulfite à représentation réduite utilisant de l'uracile n-glycosylase (ung) et de l'endonucléase iv
EP3191606B1 (fr) 2014-09-12 2020-05-27 Illumina, Inc. Procédés de détection de la présence de sous-motifs polymères par chimioluminescence
AU2015318017B2 (en) 2014-09-18 2022-02-03 Illumina, Inc. Methods and systems for analyzing nucleic acid sequencing data
EP3201355B1 (fr) 2014-09-30 2019-07-31 Illumina, Inc. Polymérases modifiées pour l'incorporation améliorée d'analogues de nucléotides
US9897791B2 (en) 2014-10-16 2018-02-20 Illumina, Inc. Optical scanning systems for in situ genetic analysis
AU2015331739B2 (en) 2014-10-17 2021-12-02 Illumina Cambridge Limited Contiguity preserving transposition
SI3212684T1 (sl) 2014-10-31 2020-04-30 Illumina Cambridge Limited Polimeri in DNK kopolimer premazi
GB201419731D0 (en) 2014-11-05 2014-12-17 Illumina Cambridge Ltd Sequencing from multiple primers to increase data rate and density
US9828627B2 (en) 2014-11-05 2017-11-28 Illumina Cambridge Limited Reducing DNA damage during sample preparation and sequencing using siderophore chelators
WO2016077350A1 (fr) 2014-11-11 2016-05-19 Illumina, Inc. Enrichissement polynucléotidique à l'aide de systèmes crispr-cas
DK3218511T3 (da) 2014-11-11 2020-07-20 Illumina Cambridge Ltd Fremgangsmåder og arrays til frembringelse og sekventering af monoklonale klynger af nukleinsyre
EP3882356A1 (fr) 2014-12-15 2021-09-22 Illumina, Inc. Compositions et procédés de positionnement de molécules individuelles sur un substrat
US10302972B2 (en) 2015-01-23 2019-05-28 Pacific Biosciences Of California, Inc. Waveguide transmission
WO2016126941A1 (fr) 2015-02-04 2016-08-11 Pacific Biosciences Of California, Inc. Réactifs fluorescents protégés multimères
SG11201706504RA (en) 2015-02-10 2017-09-28 Illumina Inc Methods and compositions for analyzing cellular components
EP3271762B1 (fr) 2015-03-16 2023-01-04 Pacific Biosciences of California, Inc. Système d'analyse comprenant dispositifs intégrés et systèmes pour couplage optique par espace libre
EP3271073B1 (fr) 2015-03-20 2019-06-12 Illumina, Inc. Cartouche fluidique pour une utilisation dans la position verticale ou sensiblement verticale
CN107810404B (zh) 2015-03-24 2020-11-10 伊鲁米那股份有限公司 用于对样品成像用于生物或化学分析的方法、载体组件和系统
ES2846730T3 (es) 2015-03-31 2021-07-29 Illumina Cambridge Ltd Concatemerización en superficie de moldes
EP4282977B1 (fr) 2015-04-10 2024-06-05 10x Genomics Sweden AB Analyse de plusieurs acides nucléiques spatialement différenciés de spécimens biologiques
CN112229834B (zh) 2015-04-14 2024-11-26 亿明达股份有限公司 用于改进对光发射的检测的结构化基底及涉及其的方法
US10844428B2 (en) 2015-04-28 2020-11-24 Illumina, Inc. Error suppression in sequenced DNA fragments using redundant reads with unique molecular indices (UMIS)
CA2983882C (fr) 2015-05-07 2023-10-03 Scott E. Helgesen Architecture pipeline de multiprocesseur
EP4190912A1 (fr) 2015-05-11 2023-06-07 Illumina, Inc. Plate-forme pour la découverte et l'analyse d'agents thérapeutiques
CN107873060B (zh) 2015-05-12 2021-10-15 亿明达股份有限公司 用于对核酸测序的场效应装置和方法
US10640809B2 (en) 2015-05-29 2020-05-05 Epicentre Technologies Corporation Methods of analyzing nucleic acids
EP3302804B1 (fr) 2015-05-29 2022-07-13 Illumina, Inc. Porte-échantillon et système de dosage permettant de provoquer des réactions indiquées
ES2789349T3 (es) 2015-05-29 2020-10-26 Illumina Cambridge Ltd Utilización mejorada de cebadores de superficie en grupos
KR102828319B1 (ko) 2015-06-03 2025-07-02 일루미나, 인코포레이티드 나노기공에 인접한 중합효소에 고정된 테터를 이용하여 폴리뉴클레오타이드를 서열결정하기 위한 조성물, 시스템 및 방법
US10365434B2 (en) 2015-06-12 2019-07-30 Pacific Biosciences Of California, Inc. Integrated target waveguide devices and systems for optical coupling
US10889861B2 (en) 2015-06-17 2021-01-12 The Translational Genomics Research Institute Systems and methods for obtaining biological molecules from a sample
US12157910B2 (en) 2015-07-06 2024-12-03 Illumina Cambridge Limited Sample preparation for nucleic acid amplification
WO2017007753A1 (fr) 2015-07-07 2017-01-12 Illumina, Inc. Formation de motifs en surface sélective par nanoimpression
US20180207920A1 (en) 2015-07-17 2018-07-26 Illumina, Inc. Polymer sheets for sequencing applications
CA3242290A1 (fr) 2015-07-27 2017-02-02 Illumina, Inc. Cartographie spatiale d'informations de sequence d'acide nucleique
WO2017019278A1 (fr) 2015-07-30 2017-02-02 Illumina, Inc. Déblocage orthogonal de nucléotides
DK3334839T3 (da) 2015-08-14 2021-04-26 Illumina Inc Systemer og fremgangsmåder under anvendelse af magnetisk reagerende sensorer til bestemmelse af en genetisk egenskab
US10976334B2 (en) 2015-08-24 2021-04-13 Illumina, Inc. In-line pressure accumulator and flow-control system for biological or chemical assays
JP6743150B2 (ja) 2015-08-28 2020-08-19 イルミナ インコーポレイテッド 単一細胞の核酸配列分析
US10906044B2 (en) 2015-09-02 2021-02-02 Illumina Cambridge Limited Methods of improving droplet operations in fluidic systems with a filler fluid including a surface regenerative silane
US10450598B2 (en) 2015-09-11 2019-10-22 Illumina, Inc. Systems and methods for obtaining a droplet having a designated concentration of a substance-of-interest
WO2017087724A1 (fr) 2015-11-17 2017-05-26 Omniome, Inc. Procédés permettant de déterminer des profils de séquences
US10781483B2 (en) 2015-11-20 2020-09-22 Pacific Biosciences Of California, Inc. Labeled nucleotide analogs, reaction mixtures, and methods and systems for sequencing
US10676788B2 (en) 2015-11-20 2020-06-09 Pacific Biosciences Of California, Inc. Modified nucleotide reagents
CN117024498A (zh) 2015-11-20 2023-11-10 加利福尼亚太平洋生物科学股份有限公司 受保护的染料标记的试剂
EP3798321B1 (fr) 2015-12-17 2025-08-27 Illumina, Inc. Différenciation des taux de méthylation dans des échantillons biologiques complexes
WO2017123533A1 (fr) 2016-01-11 2017-07-20 Illumina, Inc. Appareil de détection à microfluoromètre, système fluidique et module bride de verrouillage de cuve à circulation
TWI734748B (zh) 2016-02-17 2021-08-01 美商太斯萊特健康股份有限公司 用於生命期成像及偵測應用之感測器及裝置
WO2017165703A1 (fr) 2016-03-24 2017-09-28 Illumina, Inc. Dispositifs et compositions à base de super-réseau photonique destinés à être utilisés en imagerie par luminescence, et leurs procédés d'utilisation
EP3377226B1 (fr) 2016-03-28 2021-02-17 Illumina, Inc. Micro-réseaux multi-plans
US11326206B2 (en) 2016-04-07 2022-05-10 Pacific Biosciences Of California, Inc. Methods of quantifying target nucleic acids and identifying sequence variants
EP3235905A1 (fr) 2016-04-20 2017-10-25 QIAGEN GmbH Procédé de génération d'une bibliothèque d'arn bicaténaire
CN109072297B (zh) 2016-04-22 2022-09-13 加利福尼亚太平洋生物科学股份有限公司 使用增强的核苷酸特异性三元复合物形成检测的核酸测序方法和系统
IL301735A (en) 2016-04-22 2023-05-01 Illumina Inc Photonic stucture-based devices and compositions for use in luminescent imaging of multiple sites within a pixel, and methods of using the same
US10597643B2 (en) 2016-04-29 2020-03-24 Omniome, Inc. Polymerases engineered to reduce nucleotide-independent DNA binding
KR102231977B1 (ko) 2016-04-29 2021-03-24 옴니옴 인코포레이티드 핵산 서열의 결정 방법
CA3021769C (fr) 2016-04-29 2021-11-23 Omniome, Inc. Procede de sequencage faisant appel a la destabilisation d'un complexe ternaire pour identifier des nucleotides apparentes
WO2017197027A1 (fr) 2016-05-11 2017-11-16 Illumina, Inc. Enrichissement et amplification de polynucléotides à l'aide de systèmes d'argonaute
SG11201809376WA (en) 2016-05-18 2018-12-28 Illumina Inc Self assembled patterning using patterned hydrophobic surfaces
ES2873723T3 (es) 2016-07-22 2021-11-03 Univ Oregon Health & Science Colecciones de genoma completo de células individuales y métodos de indexación combinatoria para prepararlas
EP4039824A1 (fr) 2016-08-15 2022-08-10 Pacific Biosciences of California, Inc. Procédé et système de séquençage d'acides nucléiques
WO2018034780A1 (fr) 2016-08-15 2018-02-22 Omniome, Inc. Procédé de séquençage destiné à l'identification et au traitement rapides de paires nucléotidiques apparentées
WO2018064116A1 (fr) 2016-09-28 2018-04-05 Illumina, Inc. Procédés et systèmes de traitement de données
MY194951A (en) 2016-10-14 2022-12-28 Illumina Inc Cartridge assembly
WO2018075785A1 (fr) 2016-10-19 2018-04-26 Illumina, Inc. Méthodes de ligature chimique d'acides nucléiques
AU2017360993A1 (en) 2016-11-16 2019-06-13 Illumina, Inc. Validation methods and systems for sequence variant calls
GB201619458D0 (en) 2016-11-17 2017-01-04 Spatial Transcriptomics Ab Method for spatial tagging and analysing nucleic acids in a biological specimen
JP7228514B2 (ja) 2016-12-09 2023-02-24 ザ・ブロード・インスティテュート・インコーポレイテッド Ｃｒｉｓｐｒエフェクターシステムベースの診断法
CN118099177A (zh) 2016-12-22 2024-05-28 宽腾矽公司 具有直接合并像素的整合式光电侦测器
IL267444B2 (en) 2016-12-22 2024-12-01 Illumina Inc Arrays including a resin film and a patterned polymer layer
MY203439A (en) 2016-12-22 2024-06-28 Illumina Inc Array including sequencing primer and non-sequencing entity
CN110325651B (zh) 2016-12-22 2024-03-15 伊鲁米那股份有限公司 具有质量控制示踪剂的阵列
US11248254B2 (en) 2016-12-30 2022-02-15 Omniome, Inc. Method and system employing distinguishable polymerases for detecting ternary complexes and identifying cognate nucleotides
GB201704754D0 (en) 2017-01-05 2017-05-10 Illumina Inc Kinetic exclusion amplification of nucleic acid libraries
WO2018128544A1 (fr) 2017-01-06 2018-07-12 Agendia N.V. Biomarqueurs pour la sélection de groupes de patients, et leurs utilisations
CN110383386B (zh) 2017-01-06 2023-06-13 伊鲁米那股份有限公司 移相校正
AU2018208462B2 (en) 2017-01-10 2021-07-29 Pacific Biosciences Of California, Inc. Polymerases engineered to reduce nucleotide-independent DNA binding
JP6806909B2 (ja) 2017-01-17 2021-01-06 イルミナ インコーポレイテッド 腫瘍形成性スプライスバリアントの判定
CA3050247A1 (fr) 2017-01-18 2018-07-26 Illumina, Inc. Procedes et systemes de generation et de correction d'erreur d'ensembles d'indices moleculaires uniques ayant des longueurs moleculaires heterogenes
CN110177884B (zh) 2017-01-20 2021-08-06 欧姆尼欧美公司 通过聚合酶结合进行的基因分型
AU2018210944B2 (en) 2017-01-20 2021-07-22 Pacific Biosciences Of California, Inc. Process for cognate nucleotide detection in a nucleic acid sequencing workflow
EP3571313B1 (fr) 2017-01-20 2020-12-23 Omniome, Inc. Capture spécifique d'un allèle d'acides nucléiques
GB201701689D0 (en) 2017-02-01 2017-03-15 Illumia Inc System and method with fiducials of non-closed shapes
CN109414673B (zh) 2017-02-01 2021-09-07 伊鲁米那股份有限公司 具有响应于多个激发频率的基准的系统和方法
GB201701688D0 (en) 2017-02-01 2017-03-15 Illumia Inc System and method with fiducials in non-recliner layouts
GB201701686D0 (en) 2017-02-01 2017-03-15 Illunina Inc System & method with fiducials having offset layouts
WO2018152162A1 (fr) 2017-02-15 2018-08-23 Omniome, Inc. Distinction des séquences par détection de dissociation de polymérase
WO2018151601A1 (fr) 2017-02-17 2018-08-23 Stichting Vumc Diagnostic et sélection de thérapie améliorés par l'intelligence en essaim pour le cancer à l'aide de plaquettes éduquées contre les tumeurs
IL263118B2 (en) 2017-02-21 2023-12-01 Illumina Inc Tegumentation using fixed transpososomes with linkers
US11021740B2 (en) 2017-03-15 2021-06-01 The Broad Institute, Inc. Devices for CRISPR effector system based diagnostics
US11104937B2 (en) 2017-03-15 2021-08-31 The Broad Institute, Inc. CRISPR effector system based diagnostics
WO2018170340A1 (fr) 2017-03-15 2018-09-20 The Broad Institute, Inc. Diagnostics basés sur un système effecteur crispr pour la détection de virus
US11174515B2 (en) 2017-03-15 2021-11-16 The Broad Institute, Inc. CRISPR effector system based diagnostics
SG11201908680YA (en) 2017-03-20 2019-10-30 Illumina Inc Methods and compositions for preparing nucleic acid libraries
EP4053294A1 (fr) 2017-03-24 2022-09-07 Life Technologies Corporation Adaptateurs polynucléotidiques et procédés d'utilisation de ces derniers
US10737267B2 (en) 2017-04-04 2020-08-11 Omniome, Inc. Fluidic apparatus and methods useful for chemical and biological reactions
US10934584B2 (en) 2017-04-23 2021-03-02 Illumina, Inc. Compositions and methods for improving sample identification in indexed nucleic acid libraries
CN110785492B (zh) 2017-04-23 2024-04-30 伊鲁米纳剑桥有限公司 用于改进编索引的核酸文库中的样品鉴定的组合物和方法
SG11201909918XA (en) 2017-04-23 2019-11-28 Illumina Cambridge Ltd Compositions and methods for improving sample identification in indexed nucleic acid libraries
US9951385B1 (en) 2017-04-25 2018-04-24 Omniome, Inc. Methods and apparatus that increase sequencing-by-binding efficiency
US10161003B2 (en) 2017-04-25 2018-12-25 Omniome, Inc. Methods and apparatus that increase sequencing-by-binding efficiency
WO2018204423A1 (fr) 2017-05-01 2018-11-08 Illumina, Inc. Séquences index optimales pour séquençage multiplex massivement parallèle
EP4443436A3 (fr) 2017-05-08 2024-12-25 Illumina, Inc. Adaptateurs courts universels pour l'indexation de polynucléotides
DK3981884T3 (da) 2017-06-07 2023-09-04 Univ Oregon Health & Science Enkeltcelle-helgenombiblioteker til methyleringssekventering
CN111032882A (zh) 2017-06-20 2020-04-17 伊鲁米那股份有限公司 解决扩增反应中低效的方法和组合物
WO2019018366A1 (fr) 2017-07-18 2019-01-24 Omniome, Inc. Procédé de modification chimique de surfaces plastiques
TW201915473A (zh) 2017-07-24 2019-04-16 美商寬騰矽公司 手持，大量平行之生物光電裝置
EP3662482A1 (fr) 2017-07-31 2020-06-10 Illumina Inc. Système de séquençage comportant une agrégation d'échantillons biologiques multiplexés
EP3662084B1 (fr) 2017-08-01 2024-10-30 Illumina, Inc. Billes d'hydrogel pour le séquençage nucléotidique
PL3663407T3 (pl) 2017-08-01 2023-05-08 Mgi Tech Co., Ltd. Sposób sekwencjonowania kwasów nukleinowych
WO2019023924A1 (fr) 2017-08-01 2019-02-07 Helitec Limited Procédés d'enrichissement et de détermination de séquences nucléotidiques cibles
EP4446437A3 (fr) 2017-08-01 2025-01-01 Illumina, Inc. Indexation spatiale de matériel génétique et préparation de bibliothèque à l'aide de billes d'hydrogel et de cellules d'écoulement
WO2019035897A1 (fr) 2017-08-15 2019-02-21 Omniome, Inc. Appareil de balayage et procédés utiles pour la détection d'analytes chimiques ou biologiques
US11447818B2 (en) 2017-09-15 2022-09-20 Illumina, Inc. Universal short adapters with variable length non-random unique molecular identifiers
CA3066775A1 (fr) 2017-10-16 2019-04-25 Illumina, Inc. Techniques basees sur l'apprentissage profond d'apprentissage de reseaux neuronaux a convolution profonde
US11488009B2 (en) 2017-10-16 2022-11-01 Illumina, Inc. Deep learning-based splice site classification
AU2018353136B2 (en) 2017-10-19 2022-05-19 Pacific Biosciences Of California, Inc. Simultaneous background reduction and complex stabilization in binding assay workflows
CN110832305A (zh) 2018-01-08 2020-02-21 因美纳有限公司 利用基于半导体的检测的高通量测序系统和设备
US11378544B2 (en) 2018-01-08 2022-07-05 Illumina, Inc. High-throughput sequencing with semiconductor-based detection
WO2019140402A1 (fr) 2018-01-15 2019-07-18 Illumina, Inc. Classificateur de variants basé sur un apprentissage profond
BR112020015093A2 (pt) 2018-01-29 2020-12-08 The Broad Institute, Inc. Diagnóstico baseado no sistema de efetor crispr
KR20200139671A (ko) 2018-02-06 2020-12-14 옴니옴 인코포레이티드 핵산 프라이머 연장을 위한 조성물 및 기술
KR102773714B1 (ko) 2018-02-13 2025-02-27 일루미나, 인코포레이티드 하이드로겔 비드를 사용한 dna 서열분석
KR102330071B1 (ko) 2018-02-16 2021-11-23 일루미나, 인코포레이티드 전하-태그 뉴클레오타이드 및 이의 사용 방법
BR112019027603A2 (pt) 2018-03-09 2020-11-10 Illumina, Inc. sequenciamento estocástico generalizado de super-resolução
AU2019248635B2 (en) 2018-04-02 2022-01-27 Illumina, Inc. Compositions and methods for making controls for sequence-based genetic testing
WO2019200338A1 (fr) 2018-04-12 2019-10-17 Illumina, Inc. Classificateur de variantes basé sur des réseaux neuronaux profonds
AU2019255987A1 (en) 2018-04-19 2020-12-10 Pacific Biosciences Of California, Inc. Improving accuracy of base calls in nucleic acid sequencing methods
JP7511344B2 (ja) 2018-04-20 2024-07-05 イルミナ インコーポレイテッド 単一細胞を封入する方法、封入された細胞およびその使用
WO2019209426A1 (fr) 2018-04-26 2019-10-31 Omniome, Inc. Procédés et compositions destinés à stabiliser des complexes acide nucléique-nucléotide-polymérase
IL271446B2 (en) 2018-05-15 2025-04-01 Illumina Inc Compositions and methods for chemical removal and deprotection of surface-bound oligonucleotides
CN119955902A (zh) 2018-05-17 2025-05-09 伊鲁米纳公司 具有减少的扩增偏倚的高通量单细胞测序
KR102605749B1 (ko) 2018-05-25 2023-11-23 일루미나, 인코포레이티드 자간전증에 특이적인 순환 rna 시그니처
US11180794B2 (en) 2018-05-31 2021-11-23 Omniome, Inc. Methods and compositions for capping nucleic acids
WO2019231568A1 (fr) 2018-05-31 2019-12-05 Omniome, Inc. Augmentation du rapport signal/bruit lors du séquençage d'acides nucléiques
SG11202000905PA (en) 2018-06-04 2020-02-27 Illumina Inc High-throughput single-cell transcriptome libraries and methods of making and of using
US11391626B2 (en) 2018-06-22 2022-07-19 Quantum-Si Incorporated Integrated photodetector with charge storage bin of varied detection time
US12073922B2 (en) 2018-07-11 2024-08-27 Illumina, Inc. Deep learning-based framework for identifying sequence patterns that cause sequence-specific errors (SSEs)
EP3826762A1 (fr) 2018-07-23 2021-06-02 DNA Script Synthèse enzymatique massivement parallèle de brins d'acides nucléiques
WO2020023362A1 (fr) 2018-07-24 2020-01-30 Omniome, Inc. Formation en série d'espèces de complexe ternaire
WO2020022891A2 (fr) 2018-07-26 2020-01-30 Stichting Vumc Biomarqueurs pour fibrillation auriculaire
EP3833761A1 (fr) 2018-08-07 2021-06-16 The Broad Institute, Inc. Nouveaux systèmes et enzymes cas12b
BR112021002779A2 (pt) 2018-08-15 2021-05-04 Illumina, Inc. composições e métodos para melhorar o enriquecimento de bibliotecas
US11519033B2 (en) 2018-08-28 2022-12-06 10X Genomics, Inc. Method for transposase-mediated spatial tagging and analyzing genomic DNA in a biological sample
GB2574746B (en) 2018-09-17 2020-06-03 Omniome Inc Engineered polymerases for improved sequencing
US12467925B2 (en) 2018-10-03 2025-11-11 The Broad Institute, Inc. CRISPR effector system based diagnostics for hemorrhagic fever detection
NZ759665A (en) 2018-10-15 2022-07-01 Illumina Inc Deep learning-based techniques for pre-training deep convolutional neural networks
SG11202102703VA (en) 2018-10-26 2021-04-29 Illumina Inc Modulating polymer beads for dna processing
IL299237B2 (en) 2018-10-31 2024-12-01 Illumina Inc Polymerases, compositions, and methods of use
JP7332235B2 (ja) 2018-11-07 2023-08-23 イージーアイ テック （シェン チェン） カンパニー， リミテッド ポリヌクレオチドを配列決定する方法
JP2022513031A (ja) 2018-11-14 2022-02-07 ディーエヌエー スクリプト 末端デオキシヌクレオチジルトランスフェラーゼバリアントおよびその使用
US20240011086A1 (en) 2018-11-15 2024-01-11 Omniome, Inc. Electronic detection of nucleic acid structure
NL2022043B1 (en) 2018-11-21 2020-06-03 Akershus Univ Hf Tagmentation-Associated Multiplex PCR Enrichment Sequencing
KR20210098432A (ko) 2018-11-30 2021-08-10 일루미나, 인코포레이티드 단일 검정을 이용한 다수의 분석물의 분석
WO2020117653A1 (fr) 2018-12-04 2020-06-11 Omniome, Inc. Fluides à phase mixte pour séquençage d'acide nucléique et autres essais analytiques
KR20210098844A (ko) 2018-12-05 2021-08-11 일루미나, 인코포레이티드 중합효소, 조성물, 및 사용 방법
SG11202012762WA (en) 2018-12-05 2021-01-28 Illumina Cambridge Ltd Methods and compositions for cluster generation by bridge amplification
WO2020123311A2 (fr) 2018-12-10 2020-06-18 10X Genomics, Inc. Résolution de réseaux spatiaux à l'aide d'une déconvolution
CN113423841A (zh) 2018-12-13 2021-09-21 Dna斯克瑞普特公司 细胞和生物分子上的直接寡核苷酸合成
GB201820341D0 (en) 2018-12-13 2019-01-30 10X Genomics Inc Method for transposase-mediated spatial tagging and analysing genomic DNA in a biological specimen
GB201820300D0 (en) 2018-12-13 2019-01-30 10X Genomics Inc Method for spatial tagging and analysing genomic DNA in a biological specimen
CN112639126A (zh) 2018-12-14 2021-04-09 伊卢米纳剑桥有限公司 降低测序期间用未标记的核苷酸的定相
ES3032918T3 (en) 2018-12-17 2025-07-28 Illumina Cambridge Ltd Method of polynucleotide sequencing
CN112654714B (zh) 2018-12-17 2024-10-11 伊卢米纳剑桥有限公司 用于测序的引物寡核苷酸
KR102872035B1 (ko) 2018-12-18 2025-10-16 일루미나 케임브리지 리미티드 단일 표면 프라이머를 사용하는 페어드 엔드 서열분석을 위한 방법 및 조성물
WO2020132103A1 (fr) 2018-12-19 2020-06-25 Illumina, Inc. Procédés d'amélioration de la priorité de la clonalité d'un ensemble de polynucléotides
WO2020131759A1 (fr) 2018-12-19 2020-06-25 Roche Diagnostics Gmbh Nucléotides protégés en 3'
CN113227348B (zh) 2018-12-20 2025-05-27 加利福尼亚太平洋生物科学股份有限公司 用于分析核酸和其他分析物的温度控制
US11293061B2 (en) 2018-12-26 2022-04-05 Illumina Cambridge Limited Sequencing methods using nucleotides with 3′ AOM blocking group
JP7491934B2 (ja) 2019-01-03 2024-05-28 ディーエヌエー スクリプト オリゴヌクレオチドセットのワンポット合成
US11926867B2 (en) 2019-01-06 2024-03-12 10X Genomics, Inc. Generating capture probes for spatial analysis
US11649485B2 (en) 2019-01-06 2023-05-16 10X Genomics, Inc. Generating capture probes for spatial analysis
IL281664B2 (en) 2019-01-11 2024-07-01 Illumina Cambridge Ltd Complex surface-bound transposome complexes
EP3924500A1 (fr) 2019-02-12 2021-12-22 DNA Script Clivage efficace de produit dans la synthèse enzymatique sans matrice de polynucléotides
WO2020167574A1 (fr) 2019-02-14 2020-08-20 Omniome, Inc. Atténuation d'impacts défavorables de systèmes de détection sur des acides nucléiques et d'autres analytes biologiques
WO2020172197A1 (fr) 2019-02-19 2020-08-27 Ultima Genomics, Inc. Lieurs et procédés de détection et de séquençage optiques
US11680950B2 (en) 2019-02-20 2023-06-20 Pacific Biosciences Of California, Inc. Scanning apparatus and methods for detecting chemical and biological analytes
WO2020180778A1 (fr) 2019-03-01 2020-09-10 Illumina, Inc. Banques de noyaux uniques et à cellule unique à haut rendement et leurs procédés de production et d'utilisation
US11347965B2 (en) 2019-03-21 2022-05-31 Illumina, Inc. Training data generation for artificial intelligence-based sequencing
NL2023310B1 (en) 2019-03-21 2020-09-28 Illumina Inc Training data generation for artificial intelligence-based sequencing
NL2023316B1 (en) 2019-03-21 2020-09-28 Illumina Inc Artificial intelligence-based sequencing
NL2023312B1 (en) 2019-03-21 2020-09-28 Illumina Inc Artificial intelligence-based base calling
NL2023311B9 (en) 2019-03-21 2021-03-12 Illumina Inc Artificial intelligence-based generation of sequencing metadata
NL2023314B1 (en) 2019-03-21 2020-09-28 Illumina Inc Artificial intelligence-based quality scoring
WO2020191387A1 (fr) 2019-03-21 2020-09-24 Illumina, Inc. Appel de base à base d'intelligence artificielle
US11210554B2 (en) 2019-03-21 2021-12-28 Illumina, Inc. Artificial intelligence-based generation of sequencing metadata
AU2019445584B2 (en) 2019-05-15 2024-09-05 Qingdao MGI Tech Co., Ltd. Single-channel sequencing method based on self-luminescence
US11423306B2 (en) 2019-05-16 2022-08-23 Illumina, Inc. Systems and devices for characterization and performance analysis of pixel-based sequencing
US11593649B2 (en) 2019-05-16 2023-02-28 Illumina, Inc. Base calling using convolutions
WO2020243579A1 (fr) 2019-05-30 2020-12-03 10X Genomics, Inc. Procédés de détection de l'hétérogénéité spatiale d'un échantillon biologique
WO2020252186A1 (fr) 2019-06-11 2020-12-17 Omniome, Inc. Détection de mise au point étalonnée
CN113226519B (zh) 2019-07-12 2024-10-01 Illumina剑桥有限公司 使用电泳制备核酸文库
CA3125241A1 (fr) 2019-07-12 2021-01-21 Illumina Cambridge Limited Compositions et procedes de preparation de bibliotheques de sequencage d'acide nucleique a l'aide de crispr/cas9 immobilise sur un support solide
US11377655B2 (en) 2019-07-16 2022-07-05 Pacific Biosciences Of California, Inc. Synthetic nucleic acids having non-natural structures
US10656368B1 (en) 2019-07-24 2020-05-19 Omniome, Inc. Method and system for biological imaging using a wide field objective lens
AU2020319829A1 (en) 2019-07-30 2022-02-24 Dna Script Template-free enzymatic synthesis of polynucleotides using poly(A) and poly(U) polymerases
JP2022543568A (ja) 2019-08-01 2022-10-13 ディーエヌエー スクリプト ポリヌクレオチドの鋳型なしの酵素による合成における長い配列の収量の増加
JP7523526B2 (ja) 2019-08-20 2024-07-26 チンタオ エムジーアイ テック カンパニー リミテッド 発光標識及び二次発光信号の光信号速度論に基づくポリヌクレオチドの配列決定方法
JP2022547918A (ja) 2019-09-09 2022-11-16 ディーエヌエー スクリプト 光開裂性結合を用いる鋳型なし酵素的ポリヌクレオチド合成
US11180520B2 (en) 2019-09-10 2021-11-23 Omniome, Inc. Reversible modifications of nucleotides
WO2021050962A1 (fr) 2019-09-11 2021-03-18 The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Détection et classification de cancers
US20220403436A1 (en) 2019-09-23 2022-12-22 Dna Script Increasing Long-Sequence Yields in Template-Free Enzymatic Synthesis of Polynucleotides
CN118186063A (zh) 2019-10-18 2024-06-14 加利福尼亚太平洋生物科学股份有限公司 用于对核酸加帽的方法和组合物
EP4025711A2 (fr) 2019-11-08 2022-07-13 10X Genomics, Inc. Amélioration de la spécificité de la liaison d'un analyte
EP4055185A1 (fr) 2019-11-08 2022-09-14 10X Genomics, Inc. Agents de capture d'analytes marqués spatialement pour le multiplexage d'analytes
WO2021092431A1 (fr) 2019-11-08 2021-05-14 Omniome, Inc. Polymérases modifiées pour un séquençage amélioré par liaison
CA3157418A1 (fr) 2019-11-13 2021-05-20 Dna Script Synthese enzymatique sans matrice de polynucleotides a efficacite elevee
EP4010495A1 (fr) 2019-11-22 2022-06-15 Illumina, Inc. Signatures d'arn circulants spécifiques de la prééclampsie
DE202019106694U1 (de) 2019-12-02 2020-03-19 Omniome, Inc. System zur Sequenzierung von Nukleinsäuren in Fluidschaum
DE202019106695U1 (de) 2019-12-02 2020-03-19 Omniome, Inc. System zur Sequenzierung von Nukleinsäuren in Fluidschaum
EP4010489A1 (fr) 2019-12-04 2022-06-15 Illumina, Inc. Préparation de bibliothèques de séquençage d'adn pour la détection d'agents pathogènes d'adn dans le plasma
WO2021118349A1 (fr) 2019-12-10 2021-06-17 Prinses Máxima Centrum Voor Kinderoncologie B.V. Procédés de typage de tumeurs de cellules germinales
IL293820A (en) 2019-12-12 2022-08-01 Dna Script Chimeric-terminal deoxynucleotidyltransferases for template-independent enzymatic synthesis of polynucleotides
WO2021122539A1 (fr) 2019-12-16 2021-06-24 Dna Script Synthèse de polynucléotides enzymatiques sans matrice à l'aide de variants de désoxynucléotidyltransférases terminales sans dismutation
CN114829368B (zh) 2019-12-18 2024-06-04 豪夫迈·罗氏有限公司 通过使用连续标记方案的合成进行测序的方法
EP3927824A2 (fr) 2019-12-19 2021-12-29 Illumina, Inc. Banques de cellules uniques à haut débit et leurs procédés de production et d'utilisation
CN115135984B (zh) 2019-12-23 2025-12-23 10X基因组学有限公司 可逆固定试剂及其使用方法
US12241890B2 (en) 2019-12-23 2025-03-04 10X Genomics, Inc. Methods for generating barcoded nucleic acid molecules using fixed cells
DK3891300T3 (da) 2019-12-23 2023-05-22 10X Genomics Inc Fremgangsmåder til rumlig analyse ved anvendelse af rna-template-ligering
US12365942B2 (en) 2020-01-13 2025-07-22 10X Genomics, Inc. Methods of decreasing background on a spatial array
WO2021146443A1 (fr) * 2020-01-14 2021-07-22 Pacific Biosciences Of California, Inc. Cartouches de séquençage d'acide nucléique, dispositifs conditionnés, et systèmes
US12405264B2 (en) 2020-01-17 2025-09-02 10X Genomics, Inc. Electrophoretic system and method for analyte capture
US11702693B2 (en) 2020-01-21 2023-07-18 10X Genomics, Inc. Methods for printing cells and generating arrays of barcoded cells
US11732299B2 (en) 2020-01-21 2023-08-22 10X Genomics, Inc. Spatial assays with perturbed cells
US20210230681A1 (en) 2020-01-24 2021-07-29 10X Genomics, Inc. Methods for spatial analysis using proximity ligation
US11821035B1 (en) 2020-01-29 2023-11-21 10X Genomics, Inc. Compositions and methods of making gene expression libraries
US12076701B2 (en) 2020-01-31 2024-09-03 10X Genomics, Inc. Capturing oligonucleotides in spatial transcriptomics
US12110541B2 (en) 2020-02-03 2024-10-08 10X Genomics, Inc. Methods for preparing high-resolution spatial arrays
US11898205B2 (en) 2020-02-03 2024-02-13 10X Genomics, Inc. Increasing capture efficiency of spatial assays
CN115243792A (zh) 2020-02-04 2022-10-25 加利福尼亚太平洋生物科学股份有限公司 流通池及其制造和使用方法
US11732300B2 (en) 2020-02-05 2023-08-22 10X Genomics, Inc. Increasing efficiency of spatial analysis in a biological sample
US12129516B2 (en) 2020-02-07 2024-10-29 10X Genomics, Inc. Quantitative and automated permeabilization performance evaluation for spatial transcriptomics
US11835462B2 (en) 2020-02-11 2023-12-05 10X Genomics, Inc. Methods and compositions for partitioning a biological sample
US12281357B1 (en) 2020-02-14 2025-04-22 10X Genomics, Inc. In situ spatial barcoding
US12399123B1 (en) 2020-02-14 2025-08-26 10X Genomics, Inc. Spatial targeting of analytes
US12535420B2 (en) 2020-02-18 2026-01-27 Pacific Biosciences Of California, Inc. Highly multiplexed nucleic acid sequencing systems
US11807851B1 (en) 2020-02-18 2023-11-07 Ultima Genomics, Inc. Modified polynucleotides and uses thereof
BR112022016415A2 (pt) 2020-02-20 2022-10-11 Illumina Inc Múltiplas chamadas de base à base de inteligência artificial
US20210265016A1 (en) 2020-02-20 2021-08-26 Illumina, Inc. Data Compression for Artificial Intelligence-Based Base Calling
US20210265015A1 (en) 2020-02-20 2021-08-26 Illumina, Inc. Hardware Execution and Acceleration of Artificial Intelligence-Based Base Caller
US12354008B2 (en) 2020-02-20 2025-07-08 Illumina, Inc. Knowledge distillation and gradient pruning-based compression of artificial intelligence-based base caller
US11891654B2 (en) 2020-02-24 2024-02-06 10X Genomics, Inc. Methods of making gene expression libraries
CN115151652B (zh) 2020-02-25 2025-02-07 Dna斯克瑞普特公司 用于酶促合成多核苷酸的方法和设备
US11926863B1 (en) 2020-02-27 2024-03-12 10X Genomics, Inc. Solid state single cell method for analyzing fixed biological cells
AU2021230282A1 (en) 2020-03-03 2022-09-22 Pacific Biosciences Of California, Inc. Methods and compositions for sequencing double stranded nucleic acids
US11768175B1 (en) 2020-03-04 2023-09-26 10X Genomics, Inc. Electrophoretic methods for spatial analysis
EP4139446A1 (fr) 2020-04-20 2023-03-01 DNA Script Variants de désoxynucléotidyl transférase et utilisations correspondantes
ES3014967T3 (en) 2020-04-22 2025-04-28 10X Genomics Inc Methods for spatial analysis using targeted rna depletion
EP4143344A1 (fr) 2020-04-27 2023-03-08 Agendia N.V. Traitement du cancer du sein her2 négatif, mammaprint high risk 2
US11188778B1 (en) 2020-05-05 2021-11-30 Illumina, Inc. Equalization-based image processing and spatial crosstalk attenuator
US20230183798A1 (en) 2020-05-05 2023-06-15 Pacific Biosciences Of California, Inc. Compositions and methods for modifying polymerase-nucleic acid complexes
EP4150065A2 (fr) 2020-05-12 2023-03-22 Illumina Inc Génération d'acides nucléiques avec des bases modifiées au moyen de désoxynucléotidyl transférase terminale recombinante
US12416603B2 (en) 2020-05-19 2025-09-16 10X Genomics, Inc. Electrophoresis cassettes and instrumentation
EP4153776B1 (fr) 2020-05-22 2025-03-05 10X Genomics, Inc. Analyse spatiale pour détecter des variants de séquence
EP4414459B1 (fr) 2020-05-22 2025-09-03 10X Genomics, Inc. Mesure spatio-temporelle simultanée de l'expression génique et de l'activité cellulaire
WO2021242834A1 (fr) 2020-05-26 2021-12-02 10X Genomics, Inc. Procédé de réinitialisation d'un réseau
EP4158054B1 (fr) 2020-06-02 2025-04-16 10X Genomics, Inc. Trancriptomique spatiale pour les récepteurs d'antigènes
EP4025692A2 (fr) 2020-06-02 2022-07-13 10X Genomics, Inc. Procédés de banques d'acides nucléiques
US12265079B1 (en) 2020-06-02 2025-04-01 10X Genomics, Inc. Systems and methods for detecting analytes from captured single biological particles
US12031177B1 (en) 2020-06-04 2024-07-09 10X Genomics, Inc. Methods of enhancing spatial resolution of transcripts
EP4162075A4 (fr) 2020-06-08 2024-06-26 The Broad Institute, Inc. Indexation combinatoire à cellule unique, à partir d'acides nucléiques amplifiés
EP4421186B1 (fr) 2020-06-08 2025-08-13 10X Genomics, Inc. Procédés de détermination d'une marge chirurgicale et procédés d'utilisation de ceux-ci
WO2021252617A1 (fr) 2020-06-09 2021-12-16 Illumina, Inc. Procédés pour augmenter le rendement de bibliothèques de séquençage
WO2021252747A1 (fr) 2020-06-10 2021-12-16 1Ox Genomics, Inc. Procédés de distribution de fluide
WO2021252591A1 (fr) 2020-06-10 2021-12-16 10X Genomics, Inc. Procédés de détermination d'un emplacement d'un analyte dans un échantillon biologique
US12435363B1 (en) 2020-06-10 2025-10-07 10X Genomics, Inc. Materials and methods for spatial transcriptomics
KR20230024399A (ko) 2020-06-16 2023-02-20 디엔에이 스크립트 효소적 폴리뉴클레오티드 합성을 위한 시스템, 장치 및 키트
US12522877B2 (en) 2020-06-17 2026-01-13 The Translational Genomics Research Institute Early detection of drug-resistant mycobacterium tuberculosis
EP4168421A1 (fr) 2020-06-22 2023-04-26 Illumina Cambridge Limited Nucléosides et nucléotides avec groupe de blocage acétal 3
EP4450639B1 (fr) 2020-06-25 2025-10-15 10X Genomics, Inc. Analyse spatiale de la méthylation de l'adn
JP2023532231A (ja) 2020-06-30 2023-07-27 イルミナ インコーポレイテッド 傷なしｄｎａを生成するための触媒的に制御された合成による配列決定
CN115843315A (zh) 2020-07-02 2023-03-24 伊鲁米纳公司 用于校准流通池中核酸文库接种效率的方法
US12209280B1 (en) 2020-07-06 2025-01-28 10X Genomics, Inc. Methods of identifying abundance and location of an analyte in a biological sample using second strand synthesis
US11981960B1 (en) 2020-07-06 2024-05-14 10X Genomics, Inc. Spatial analysis utilizing degradable hydrogels
US11761038B1 (en) 2020-07-06 2023-09-19 10X Genomics, Inc. Methods for identifying a location of an RNA in a biological sample
WO2022010965A1 (fr) 2020-07-08 2022-01-13 Illumina, Inc. Billes utilisées en tant que supports pour transposomes
EP4182472A1 (fr) 2020-07-15 2023-05-24 DNA Script Synthèse enzymatique massivement parallèle de polynucléotides
CN116171330A (zh) 2020-08-06 2023-05-26 Illumina公司 使用小珠连接的转座体制备rna和dna测序文库
US12553898B1 (en) 2020-08-10 2026-02-17 10X Genomics, Inc. Fluorescent hybridization of antibody-oligonucleotide for multiplexing and signal amplification
IL299783A (en) 2020-08-18 2023-03-01 Illumina Inc Sequence-Specific Targeted Transposition, Selection and Sorting of Nucleic Acids
US11981958B1 (en) 2020-08-20 2024-05-14 10X Genomics, Inc. Methods for spatial analysis using DNA capture
MX2023001400A (es) 2020-09-11 2023-04-25 Illumina Cambridge Ltd Metodos para enriquecer una secuencia diana a partir de una genoteca de secuenciación usando adaptadores de horquilla.
EP4214678B1 (fr) 2020-09-18 2024-11-06 10X Genomics, Inc. Appareil de manipulation d'échantillons et procédés d'enregistrement d'images
CA3193386A1 (fr) 2020-09-22 2022-03-31 Mikhael SOSKINE Variants stabilises de desoxynucleotidyl transferase terminale tronques a l'extremite n-terminale et utilisations associees
US11926822B1 (en) 2020-09-23 2024-03-12 10X Genomics, Inc. Three-dimensional spatial analysis
CN116438319A (zh) 2020-10-21 2023-07-14 Illumina公司 包含多个插入物的测序模板以及用于提高测序通量的组合物和方法
WO2022090057A1 (fr) 2020-10-26 2022-05-05 Dna Script Nouveaux variants d'endonucléase v et leurs utilisations
US20240052391A1 (en) 2020-10-29 2024-02-15 Dna Script Enzymatic Synthesis of Polynucleotide Probes
US11827935B1 (en) 2020-11-19 2023-11-28 10X Genomics, Inc. Methods for spatial analysis using rolling circle amplification and detection probes
WO2022140028A1 (fr) 2020-12-21 2022-06-30 10X Genomics, Inc. Procédés, compositions et systèmes pour capturer des sondes et/ou des codes à barres
CA3204784A1 (fr) 2021-01-13 2022-07-21 Alex Nemiroski Structuration de surface a l'aide d'un ensemble colloidal
JP2024505508A (ja) 2021-01-29 2024-02-06 イルミナ インコーポレイテッド ポリヌクレオチドによるフローセルの播種効率を改善するための方法、組成物、及びキット
CA3208854A1 (fr) 2021-02-04 2022-08-11 Illumina, Inc. Generation de lectures longues indexees-liees sur des billes liees par transposomes
US20240043915A1 (en) 2021-02-13 2024-02-08 The General Hospital Corporation Methods and compositions for in situ macromolecule detection and uses thereof
WO2022178267A2 (fr) 2021-02-19 2022-08-25 10X Genomics, Inc. Dispositifs de support de dosage modulaires
WO2022197754A1 (fr) 2021-03-16 2022-09-22 Illumina Software, Inc. Quantification de paramètres de réseau neuronal pour appel de base
US12525320B2 (en) 2021-03-16 2026-01-13 Illumina, Inc. Neural network parameter quantization for base calling
WO2022198068A1 (fr) 2021-03-18 2022-09-22 10X Genomics, Inc. Capture multiplex de gène et expression de protéines à partir d'un échantillon biologique
US20220333178A1 (en) 2021-03-22 2022-10-20 Illumina Cambridge Limited Methods for improving nucleic acid cluster clonality
EP4314328A1 (fr) 2021-03-29 2024-02-07 Illumina, Inc. Compositions et procédés pour évaluer l'endommagement de l'adn dans une bibliothèque et normaliser une polarisation de taille d'amplicon
WO2022212269A1 (fr) 2021-03-29 2022-10-06 Illumina, Inc. Procédés améliorés de préparation de banques
MX2023010495A (es) 2021-03-30 2023-09-18 Illumina Inc Metodos mejorados de preparacion de genoteca y adn complementarios isotermicos.
BR112023019945A2 (pt) 2021-03-31 2023-11-14 Illumina Cambridge Ltd Métodos para a preparação de bibliotecas de sequenciamento por tagmentação direcional com o uso de tecnologia baseada em transposon com identificadores moleculares exclusivos para correção de erros
CN117222744A (zh) 2021-04-02 2023-12-12 Dna斯克瑞普特公司 用于酶法合成g4-易发的多核苷酸的方法和试剂盒
KR20230167028A (ko) 2021-04-02 2023-12-07 일루미나, 인코포레이티드 시퀀싱을 위한 뉴클레오티드 샘플 슬라이드 내 거품을 검출하기 위한 기계-학습 모델
EP4305196B1 (fr) 2021-04-14 2025-04-02 10X Genomics, Inc. Procédés de mesure d'une mauvaise localisation d'un analyte
US20220336057A1 (en) 2021-04-15 2022-10-20 Illumina, Inc. Efficient voxelization for deep learning
US12217829B2 (en) 2021-04-15 2025-02-04 Illumina, Inc. Artificial intelligence-based analysis of protein three-dimensional (3D) structures
US12070744B2 (en) 2021-04-22 2024-08-27 Illumina, Inc. Valve assemblies and related systems
JP2024516290A (ja) 2021-05-04 2024-04-12 パシフィック・バイオサイエンシズ・オブ・カリフォルニア・インク． 縮小されたスケールの単位セルを伴う統合された分析デバイスのアレイ
WO2022236054A1 (fr) 2021-05-06 2022-11-10 10X Genomics, Inc. Procédés pour augmenter la résolution d'une analyse spatiale
EP4334475A1 (fr) 2021-05-07 2024-03-13 Agendia N.V. Traitement endocrinien du cancer du sein à récepteurs hormonaux positifs identifié comme présentant un faible risque de récidive
EP4337786A1 (fr) 2021-05-10 2024-03-20 Pacific Biosciences of California, Inc. Ensemencement et amplification monomoléculaire sur une surface
CN117858959A (zh) 2021-05-10 2024-04-09 加利福尼亚太平洋生物科学股份有限公司 滚环扩增期间的dna扩增缓冲液补充
WO2022243480A1 (fr) 2021-05-20 2022-11-24 Illumina, Inc. Compositions et procédés de séquençage par synthèse
EP4582555A3 (fr) 2021-06-03 2025-10-22 10X Genomics, Inc. Procédés, compositions, kits et systèmes pour améliorer la capture d'analytes pour une analyse spatiale
WO2022265994A1 (fr) 2021-06-15 2022-12-22 Illumina, Inc. Fonctionnalisation de surface exempte d'hydrogel pour séquençage
EP4359555A1 (fr) 2021-06-23 2024-05-01 Illumina, Inc. Compositions, procédés, kits, cartouches et systèmes de séquençage de réactifs
WO2023278184A1 (fr) 2021-06-29 2023-01-05 Illumina, Inc. Procédés et systèmes pour corriger une diaphonie dans un éclairage émis par des sites de réaction
KR20240027608A (ko) 2021-06-29 2024-03-04 일루미나, 인코포레이티드 유기체 서열을 사용하여 훈련된 자체-학습 염기 호출자
AU2022301321A1 (en) 2021-06-29 2024-01-18 Illumina, Inc. Machine-learning model for generating confidence classifications for genomic coordinates
MX2023015504A (es) 2021-06-29 2024-01-22 Illumina Inc Metrica de la relacion se?al a ruido para determinar llamadas de bases de nucleotidos y la calidad de las llamadas de base.
US12530882B2 (en) 2021-07-01 2026-01-20 Illumina, Inc. Efficient artificial intelligence-based base calling of index sequences
US12423815B2 (en) 2021-07-13 2025-09-23 Illumina, Inc. Methods and systems for real time extraction of crosstalk in illumination emitted from reaction sites
JP2024532004A (ja) 2021-07-19 2024-09-04 イルミナ インコーポレイテッド ベースコールのための補間及び適合による強度抽出
US11455487B1 (en) 2021-10-26 2022-09-27 Illumina Software, Inc. Intensity extraction and crosstalk attenuation using interpolation and adaptation for base calling
US20230021577A1 (en) 2021-07-23 2023-01-26 Illumina Software, Inc. Machine-learning model for recalibrating nucleotide-base calls
US20230116852A1 (en) 2021-07-23 2023-04-13 Illumina, Inc. Methods for preparing substrate surface for dna sequencing
WO2023009758A1 (fr) 2021-07-28 2023-02-02 Illumina, Inc. Étalonnage de score de qualité de systèmes d'appel de bases
US12553805B2 (en) 2021-08-02 2026-02-17 10X Genomics, Inc. Methods of preserving a biological sample
KR20240035413A (ko) 2021-08-03 2024-03-15 일루미나, 인코포레이티드 다중 염기 호출자 모델을 사용한 염기 호출
US12077789B2 (en) 2021-08-14 2024-09-03 Illumina, Inc. Polymerases, compositions, and methods of use
AU2022331421A1 (en) 2021-08-17 2024-01-04 Illumina, Inc. Methods and compositions for identifying methylated cytosines
EP4344443A1 (fr) 2021-08-31 2024-04-03 Illumina, Inc. Cellule d'écoulement à résolution d'imagerie de puits améliorée
EP4196605B1 (fr) 2021-09-01 2024-12-04 10X Genomics, Inc. Procédés pour bloquer une sonde de capture sur un réseau spatial
CN115943217B (zh) 2021-09-07 2026-02-24 深圳华大智造科技股份有限公司 一种用于分析靶多核苷酸的序列的方法
CA3231118A1 (fr) 2021-09-07 2023-03-16 Meihua Gong Procede d'analyse de la sequence d'un polynucleotide cible
WO2023044229A1 (fr) 2021-09-17 2023-03-23 Illumina, Inc. Identification automatique de sources de défaillance en séquençage nucléotidique à partir de motifs d'erreur d'appel de base
US20230095961A1 (en) 2021-09-21 2023-03-30 Illumina, Inc. Graph reference genome and base-calling approach using imputed haplotypes
WO2023049215A1 (fr) 2021-09-22 2023-03-30 Illumina, Inc. Appel de base basé sur l'état compressé
US12480157B2 (en) 2021-09-30 2025-11-25 Illumina, Inc. Polynucleotide sequencing
WO2023056328A2 (fr) 2021-09-30 2023-04-06 Illumina, Inc. Supports solides et procédés d'appauvrissement et/ou d'enrichissement de fragments de bibliothèque préparés à partir de bioéchantillons
US20250236865A1 (en) 2021-10-20 2025-07-24 Illumina, Inc. Methods for capturing library dna for sequencing
EP4174189A1 (fr) 2021-10-28 2023-05-03 Volker, Leen Étiquetage des biomolécules dirigé par les enzymes
WO2023081485A1 (fr) 2021-11-08 2023-05-11 Pacific Biosciences Of California, Inc. Séquençage par étapes d'un polynucléotide à l'aide d'un mélange réactionnel homogène
WO2023086880A1 (fr) 2021-11-10 2023-05-19 10X Genomics, Inc. Procédés, compositions et kits pour déterminer l'emplacement d'un analyte dans un échantillon biologique
WO2023083997A2 (fr) 2021-11-10 2023-05-19 Dna Script Nouveau désoxynucléotidyl terminal
WO2023085932A1 (fr) 2021-11-10 2023-05-19 Omnigen B.V. Prédiction de la réponse au traitement par folfirinox chez les patients atteints de cancer
US20250002879A1 (en) 2021-11-10 2025-01-02 Dna Script NOVEL TERMINAL DEOXYNUCLEOTIDYL TRANSFERASE (TdT) VARIANTS
WO2023102118A2 (fr) 2021-12-01 2023-06-08 10X Genomics, Inc. Procédés, compositions et systèmes pour la détection améliorée d'analytes in situ et analyse spatiale
US20230343415A1 (en) 2021-12-02 2023-10-26 Illumina Software, Inc. Generating cluster-specific-signal corrections for determining nucleotide-base calls
EP4441711A1 (fr) 2021-12-20 2024-10-09 10X Genomics, Inc. Auto-test pour dispositif d'imagerie
CN117597569A (zh) 2021-12-23 2024-02-23 伊鲁米那有限公司 系统和相关的温度校准方法
WO2023122363A1 (fr) 2021-12-23 2023-06-29 Illumina Software, Inc. Résumés dynamiques d'état graphique pour séquençage de nucléotides
US20230215515A1 (en) 2021-12-23 2023-07-06 Illumina Software, Inc. Facilitating secure execution of external workflows for genomic sequencing diagnostics
US20230207050A1 (en) 2021-12-28 2023-06-29 Illumina Software, Inc. Machine learning model for recalibrating nucleotide base calls corresponding to target variants
EP4457814A1 (fr) 2021-12-29 2024-11-06 Illumina, Inc. Permutation automatique des versions de modèle d'analyse de variants pour les applications d'analyse génomique
US20240294967A1 (en) 2022-01-20 2024-09-05 Illumina, Inc. Methods of detecting methylcytosine and hydroxymethylcytosine by sequencing
BE1030246B1 (nl) 2022-02-04 2023-09-04 Leen Volker Polymeer geassisteerde biomolecule analyse
KR20240152324A (ko) 2022-02-25 2024-10-21 일루미나, 인코포레이티드 뉴클레오티드 서열분석을 위한 교정 서열
AU2023225949A1 (en) 2022-02-25 2024-01-18 Illumina, Inc. Machine-learning models for detecting and adjusting values for nucleotide methylation levels
US20230343414A1 (en) 2022-03-25 2023-10-26 Illumina, Inc. Sequence-to-sequence base calling
JP2025512975A (ja) 2022-04-07 2025-04-22 イルミナ インコーポレイテッド 改変シチジンデアミナーゼ及び使用方法
US20230340571A1 (en) 2022-04-26 2023-10-26 Illumina, Inc. Machine-learning models for selecting oligonucleotide probes for array technologies
US20230348967A1 (en) 2022-04-29 2023-11-02 Illumina Cambridge Limited Methods and systems for encapsulating lyophilised microspheres
WO2023220627A1 (fr) 2022-05-10 2023-11-16 Illumina Software, Inc. Réseau neuronal adaptatif pour séquençage de nucléotides
US20260049359A1 (en) 2022-05-19 2026-02-19 Agendia N.V. Prediction of response to immune therapy in breast cancer patients
WO2023224488A1 (fr) 2022-05-19 2023-11-23 Agendia N.V. Signature de réparation d'adn et prédiction de réponse après une cancérothérapie
US20230392207A1 (en) 2022-06-03 2023-12-07 Illumina, Inc. Circulating rna biomarkers for preeclampsia
CN119317964A (zh) 2022-06-09 2025-01-14 因美纳有限公司 碱基检出对流通池倾斜的依赖
CN119422201B (zh) 2022-06-24 2026-03-06 因美纳有限公司 通过智能鉴定候选分割组并评分来改进分割-读段比对的方法
JP2025523561A (ja) 2022-06-27 2025-07-23 イルミナ インコーポレイテッド 構造多型グラフゲノムの生成及び実装
EP4544554A1 (fr) 2022-06-27 2025-04-30 Illumina, Inc. Génotypage amélioré d'antigène leucocytaire humain (hla)
KR20250034302A (ko) 2022-06-27 2025-03-11 일루미나, 인코포레이티드 유전자형 대치 모델을 위한 가속기
WO2024015962A1 (fr) 2022-07-15 2024-01-18 Pacific Biosciences Of California, Inc. Adaptateurs en épingle à cheveux asymétriques bloqués
WO2024026356A1 (fr) 2022-07-26 2024-02-01 Illumina, Inc. Traitement multiomique à cellule unique rapide à l'aide d'un fichier exécutable
EP4595060A1 (fr) 2022-09-29 2025-08-06 Illumina, Inc. Panneau de référence de variant cible servant à imputer des variants cibles
WO2024069581A1 (fr) 2022-09-30 2024-04-04 Illumina Singapore Pte. Ltd. Complexes hélicase-cytidine désaminase et procédés d'utilisation
WO2024073043A1 (fr) 2022-09-30 2024-04-04 Illumina, Inc. Procédés d'utilisation de protéines de liaison cpg dans la cartographie de nucléotides cytosine modifiés
US20240141427A1 (en) 2022-09-30 2024-05-02 Illumina, Inc. Polymerases, compositions, and methods of use
EP4595059A1 (fr) 2022-09-30 2025-08-06 Illumina, Inc. Modèle d'apprentissage automatique pour affiner des appels de variants structuraux
WO2024073047A1 (fr) 2022-09-30 2024-04-04 Illumina, Inc. Cytidine désaminases et procédés d'utilisation dans la cartographie de nucléotides cytosine modifiés
EP4599449A1 (fr) 2022-10-05 2025-08-13 Illumina, Inc. Intégration d'appels de variants à partir de multiples pipelines de séquençage à l'aide d'une architecture d'apprentissage automatique
WO2024077202A2 (fr) 2022-10-06 2024-04-11 Illumina, Inc. Sondes pour améliorer la surveillance d'échantillons environnementaux
EP4482978A1 (fr) 2022-10-06 2025-01-01 Illumina, Inc. Sondes servant à appauvrir un petit arn non codant abondant
EP4482987A2 (fr) 2022-10-06 2025-01-01 Illumina, Inc. Sondes pour améliorer la surveillance d'échantillons de coronavirus
EP4602608A1 (fr) 2022-10-11 2025-08-20 Illumina, Inc. Détection et correction de valeurs de méthylation à partir de dosages de séquençage de méthylation
EP4605542A1 (fr) 2022-10-19 2025-08-27 DNA Script Procédés et produits pour l'élimination de polynucléotides contenant de l'uracile
EP4482979B1 (fr) 2022-11-09 2025-10-01 10X Genomics, Inc. Procédés, compositions et kits pour déterminer l'emplacement de plusieurs analytes dans un échantillon biologique
WO2024118791A1 (fr) 2022-11-30 2024-06-06 Illumina, Inc. Prédiction précise de variants à partir de données de séquençage de méthylation
EP4627113A1 (fr) 2022-11-30 2025-10-08 Illumina, Inc. Correction chimio-enzymatique de transformations d'uracile faux positives
EP4630540A1 (fr) 2022-12-05 2025-10-15 DNA Script Variants de poly (a) polymérase et leurs utilisations
WO2024129672A1 (fr) 2022-12-12 2024-06-20 The Broad Institute, Inc. Arn en circulation pour évaluer la connectivité cellule-cellule et la neuroanatomie
WO2024129969A1 (fr) 2022-12-14 2024-06-20 Illumina, Inc. Systèmes et procédés de capture et d'enrichissement de billes en grappe sur des substrats de cuve à circulation
EP4634403A1 (fr) 2022-12-16 2025-10-22 Illumina, Inc. Boranes sur supports solides
EP4638795A1 (fr) 2022-12-22 2025-10-29 Illumina, Inc. Compositions de catalyseur de métal de transition et procédés de séquençage par synthèse
US20240247311A1 (en) 2022-12-22 2024-07-25 Illumina, Inc. Palladium catalyst compositions and methods for sequencing by synthesis
CA3246559A1 (fr) 2022-12-27 2024-07-04 Illumina, Inc. Procédés de séquençage utilisant des nucléotides bloqués par allyle 3'
WO2024141628A1 (fr) 2022-12-31 2024-07-04 Dna Script Encres à viscosité variable pour l'administration par jet d'encre de réactifs enzymatiques
CN119301271A (zh) 2023-01-06 2025-01-10 因美纳有限公司 通过聚合酶减少尿嘧啶
EP4652292A1 (fr) 2023-01-16 2025-11-26 DNA Script Synthèse enzymatique d'acides nucléiques assistée par jet d'encre
CN120530202A (zh) 2023-01-26 2025-08-22 Dna斯克瑞普特公司 多核苷酸探针的酶促合成
JP2026506415A (ja) 2023-02-06 2026-02-25 イルミナ インコーポレイテッド クラスタ間光干渉の決定及び除去
EP4665873A1 (fr) 2023-02-17 2025-12-24 Illumina, Inc. Signaux d'adn acellulaire utilisés en tant que biomarqueurs de la prééclampsie
WO2024206394A1 (fr) 2023-03-30 2024-10-03 Illumina, Inc. Compositions et procédés de séquençage d'acides nucléiques
EP4706045A1 (fr) 2023-03-30 2026-03-11 Illumina, Inc. Génotypage à répétition en tandem
CN119744419A (zh) 2023-05-03 2025-04-01 因美纳有限公司 用于重新校准来自现有测序数据文件的基因型检出的机器学习模型
WO2024249200A1 (fr) 2023-05-26 2024-12-05 Illumina, Inc. Procédés de conservation de l'état de méthylation pendant le regroupement
US20240404624A1 (en) 2023-05-31 2024-12-05 Illumina, Inc. Structural variant alignment and variant calling by utilizing a structural-variant reference genome
WO2024249973A2 (fr) 2023-06-02 2024-12-05 Illumina, Inc. Liaison de gènes humains à des phénotypes cliniques à l'aide de réseaux neuronaux graphiques
WO2024254003A1 (fr) 2023-06-05 2024-12-12 Illumina, Inc. Identification et cartographie de sites de méthylation
WO2024256604A1 (fr) 2023-06-15 2024-12-19 Dna Script Addition de sels pendant la synthèse enzymatique de polynucléotides
CN121666455A (zh) 2023-06-26 2026-03-13 加利福尼亚太平洋生物科学股份有限公司 用于核酸延伸的组合物和方法
WO2025006565A1 (fr) 2023-06-27 2025-01-02 Illumina, Inc. Appel de variant avec estimation du niveau de méthylation
AU2024311047A1 (en) 2023-06-30 2025-12-11 Illumina, Inc. Using flowcell spatial coordinates to link reads for improved genome analysis
WO2025006874A1 (fr) 2023-06-30 2025-01-02 Illumina, Inc. Modèle d'apprentissage automatique pour réétalonner des appels de génotype correspondant à des variants de lignée germinale et variants de mosaïque somatique
WO2025010160A1 (fr) 2023-07-06 2025-01-09 Pacific Biosciences Of California, Inc. Procédés et compositions pour stabiliser des concatémères
GB202312147D0 (en) 2023-08-08 2023-09-20 Syndex Bio Ltd Methylation method
WO2025054389A1 (fr) 2023-09-07 2025-03-13 Illumina, Inc. Identification de la cytosine méthylée par l'utilisation de points de repère
WO2025059045A1 (fr) 2023-09-12 2025-03-20 Illumina, Inc. Systèmes et procédés de détermination de liaison de lectures de séquence sur une cellule de flux
WO2025058517A1 (fr) 2023-09-12 2025-03-20 Levels Diagnostics Holding B.V. Biomarqueurs pour typer un échantillon d'un individu à la recherche d'un carcinome hépatocellulaire
WO2025059533A1 (fr) 2023-09-13 2025-03-20 The Broad Institute, Inc. Enzymes crispr et systèmes
WO2025072368A1 (fr) 2023-09-28 2025-04-03 Illumina, Inc. Capture et libération sélective de matériau biologique
WO2025072783A1 (fr) 2023-09-28 2025-04-03 Illumina, Inc. Cytidine désaminases modifiées et méthodes d'utilisation
US20250111899A1 (en) 2023-09-29 2025-04-03 Illumina, Inc. Predicting insert lengths using primary analysis metrics
US20250111898A1 (en) 2023-09-29 2025-04-03 Illumina, Inc. Tracking and modifying cluster location on nucleotide-sample slides in real time
WO2025081064A2 (fr) 2023-10-11 2025-04-17 Illumina, Inc. Désaminase thermophile et procédés d'identification de cytosine modifiée
WO2025088024A1 (fr) 2023-10-24 2025-05-01 Dna Script Nouveaux variants de désoxynucléotidyl transférase terminale (tdt) et leurs utilisations
WO2025090883A1 (fr) 2023-10-27 2025-05-01 Illumina, Inc. Détection de variants dans des séquences nucléotidiques sur la base d'une diversité d'haplotype
WO2025106629A1 (fr) 2023-11-17 2025-05-22 Illumina, Inc. Détection de variante structurale à l'aide de lectures spatialement liées
WO2025106431A1 (fr) 2023-11-17 2025-05-22 Illumina, Inc. Détermination de variantes structurelles
WO2025117738A1 (fr) 2023-11-28 2025-06-05 Illumina, Inc. Procédés d'amélioration d'efficacité de ligature d'indice moléculaire unique
EP4567128A1 (fr) 2023-12-07 2025-06-11 Max-Delbrück-Centrum für Molekulare Medizin in der Helmholtz-Gemeinschaft Procede et moyens ameliores de detection spatiale d'acides nucleiques in situ
WO2025129074A2 (fr) 2023-12-14 2025-06-19 Illumina, Inc. Techniques d'indexation pour bibliothèques d'adn tagmentés
WO2025129133A1 (fr) 2023-12-15 2025-06-19 Illumina, Inc. Modèles de maladie résiduelle minime (mrd) pour déterminer des probabilités qu'un sujet comprenne un cancer
WO2025132627A1 (fr) 2023-12-18 2025-06-26 Dna Script Variants stabilisés d'endonucléase v hautement active pour le clivage d'adn marqué
WO2025132603A1 (fr) 2023-12-18 2025-06-26 Dna Script Nouveaux variants d'endonucléase v pour le clivage d'adn marqué
WO2025136998A1 (fr) 2023-12-18 2025-06-26 Illumina, Inc. Utilisation de modèles d'apprentissage automatique pour détecter une maladie résiduelle minimale (mrd) chez un sujet
WO2025137222A1 (fr) 2023-12-19 2025-06-26 Illumina, Inc. Dosage de détection de méthylation
WO2025137341A1 (fr) 2023-12-20 2025-06-26 Illumina, Inc. Détermination directe de mesures de rapport signal sur bruit pour une convergence accélérée lors de la détermination d'appels de base nucléotidique et d'une qualité d'appel de base
WO2025137268A1 (fr) 2023-12-20 2025-06-26 Pacific Biosciences Of California, Inc. Procédés et compositions pour pour réduire la dépendance à la cg
WO2025136105A1 (fr) 2023-12-20 2025-06-26 Stichting Amsterdam UMC Signatures microbiennes adhérentes au tissu intestinal prédictives de la réponse à l'anti-tnf-alpha dans la maladie de crohn
US20250210141A1 (en) 2023-12-21 2025-06-26 Illumina, Inc. Enhanced mapping and alignment of nucleotide reads utilizing an improved haplotype data structure with allele-variant differences
WO2025136717A1 (fr) 2023-12-22 2025-06-26 Illumina, Inc. Amélioration de la résolution de mappage à l'aide d'informations spatiales de lectures séquencées
US20250215493A1 (en) 2023-12-28 2025-07-03 Illumina, Inc. Nucleotides with enzymatically cleavable 3'-o-glycoside blocking groups for sequencing
WO2025160089A1 (fr) 2024-01-26 2025-07-31 Illumina, Inc. Construction de référence multigénome personnalisée pour une analyse de séquençage améliorée d'échantillons génomiques
WO2025174774A1 (fr) 2024-02-12 2025-08-21 Illumina, Inc. Détermination de corrections hors ligne pour des erreurs spécifiques de séquence provoquées par des séquences nucléotidiques de faible complexité
WO2025174708A1 (fr) 2024-02-13 2025-08-21 Illumina, Inc. Conception et procédé de compatibilité inter-plateformes de séquençage de cuves à circulation
WO2025184226A1 (fr) 2024-02-28 2025-09-04 Illumina, Inc. Nucléotides à coiffage terminal de phosphate
WO2025184234A1 (fr) 2024-02-28 2025-09-04 Illumina, Inc. Base de données d'haplotypes personnalisée pour mappage et alignement améliorés de lectures de nucléotides et appel de génotype amélioré
WO2025189105A1 (fr) 2024-03-08 2025-09-12 Illumina, Inc. Seuillage par taille de fragments d'adn
WO2025188906A1 (fr) 2024-03-08 2025-09-12 Illumina, Inc. Nucléotides d'adénosine modifiés
WO2025193747A1 (fr) 2024-03-12 2025-09-18 Illumina, Inc. Modèles d'apprentissage automatique pour ordonner et accélérer les tâches de séquençage ou les lames d'échantillons de nucléotides correspondantes
WO2025198469A1 (fr) 2024-03-18 2025-09-25 Agendia N.V. Prédiction de réponse à une thérapie immunitaire chez des patients atteints d'un cancer du sein triple négatif
WO2025207886A1 (fr) 2024-03-28 2025-10-02 Illumina, Inc. Kits et procédés de préparation de bibliothèque de cuves à circulation et de détection de méthylation
WO2025230914A1 (fr) 2024-04-29 2025-11-06 Illumina, Inc. Nucléotides avec lieurs auto-immolables déclenchés par des enzymes pour le séquençage par synthèse
WO2025240241A1 (fr) 2024-05-13 2025-11-20 Illumina, Inc. Modification de cycles de séquençage pendant une analyse de séquençage pour satisfaire des estimations de couverture personnalisées pour une région génomique cible
WO2025240924A1 (fr) 2024-05-17 2025-11-20 Illumina, Inc. Systèmes d'égalisation aveugle pour applications d'appel de base
WO2025250996A2 (fr) 2024-05-31 2025-12-04 Illumina, Inc. Modèles de génération et de réétalonnage d'appel pour mettre en œuvre des haplotypes de référence diploïdes personnalisés dans un appel de génotype
WO2025264836A1 (fr) 2024-06-18 2025-12-26 Illumina, Inc. Procédés pour augmenter la qualité de séquençage de régions riches en gc
WO2025264831A1 (fr) 2024-06-18 2025-12-26 Illumina, Inc. Procédés pour augmenter la qualité de séquençage de régions riches en gc
WO2026006314A1 (fr) 2024-06-28 2026-01-02 Illumina, Inc. Marquage de régions cibles avant le séquençage de nucléotides
WO2026006774A1 (fr) 2024-06-28 2026-01-02 Illumina, Inc. Cytidine désaminases modifiées et procédés d'utilisation
WO2026006771A1 (fr) 2024-06-28 2026-01-02 Illumina, Inc. Scores de filtrage de grappe
WO2026008711A2 (fr) 2024-07-02 2026-01-08 Dna Script Dispositif de filtrage et procédé de synthèse enzymatique de polynucléotides
WO2026008717A2 (fr) 2024-07-02 2026-01-08 Dna Script Procédé de synthèse enzymatique de polynucléotides
WO2026030370A1 (fr) 2024-07-30 2026-02-05 Illumina, Inc. Génération de limites de décision de valeur d'intensité adaptative pour séquençage de nucléotides

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4863849A (en) * 1985-07-18 1989-09-05 New York Medical College Automatable process for sequencing nucleotide
US4959463A (en) * 1985-10-15 1990-09-25 Genentech, Inc. Intermediates
US4962037A (en) * 1987-10-07 1990-10-09 United States Of America Method for rapid base sequencing in DNA and RNA
US4971903A (en) * 1988-03-25 1990-11-20 Edward Hyman Pyrophosphate-based method and apparatus for sequencing nucleic acids
GB8910880D0 (en) * 1989-05-11 1989-06-28 Amersham Int Plc Sequencing method

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO9106678A1 *

Also Published As

Publication number Publication date
CA2044616A1 (fr) 1991-04-27
WO1991006678A1 (fr) 1991-05-16

Similar Documents

Publication Publication Date Title
EP0450060A1 (fr) Sequen age d'adn
Metzker et al. Termination of DNA synthesis by novel 3'-modifieddeoxyribonucleoside 5'-triphosphates
US5547835A (en) DNA sequencing by mass spectrometry
US5728529A (en) Alternative dye-labeled ribonucleotides, deoxyribonucleotides, and dideoxyribonucleotides for automated DNA analysis
EP2321429B1 (fr) Procédés et kits de séquençage d'acides nucléiques
Matteucci et al. Synthesis of deoxyoligonucleotides on a polymer support
EP0640146B1 (fr) Procede de sequencage d'adn
JP3013156B2 (ja) ３’−固有の校正活性を有するｄｎａポリメラーゼの使用
US5908926A (en) 5'to 3' nucleic acid synthesis using 3'-photoremovable protecting group
US5547839A (en) Sequencing of surface immobilized polymers utilizing microflourescence detection
US6573374B1 (en) Nucleotides labelled with an infrared dye and their use in nucleic acid detection
WO2008037568A2 (fr) Terminateurs réversibles pour un séquençage efficace par synthèse
KR20010099663A (ko) 폴리뉴클레오티드의 분석 방법
Rosenthal et al. Automated sequencing of fluorescently labelled DNA by chemical degradation
AU694940C (en) DNA sequencing by mass spectrometry
JPH04503460A (ja) Ｄｎａ配列決定
Caruthers Sequencing and Synthesis of Nucleic Acids
Narang The Total Synthesis
Lowe ACID REPEAT SEQUENCES BY $8 $8 2i DISCONTINUOUS PRIMER EXTENSION

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): DE FR GB IT NL

17P Request for examination filed

Effective date: 19910925

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN

18D Application deemed to be withdrawn

Effective date: 19940501