DK2964624T3 - Rhodaminforbindelser og anvendelse heraf som fluorescerende mærker - Google Patents
Rhodaminforbindelser og anvendelse heraf som fluorescerende mærker Download PDFInfo
- Publication number
- DK2964624T3 DK2964624T3 DK13709182.3T DK13709182T DK2964624T3 DK 2964624 T3 DK2964624 T3 DK 2964624T3 DK 13709182 T DK13709182 T DK 13709182T DK 2964624 T3 DK2964624 T3 DK 2964624T3
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- Denmark
- Prior art keywords
- alkyl
- group
- substituted
- nucleotide
- dye
- Prior art date
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- PYWVYCXTNDRMGF-UHFFFAOYSA-N rhodamine B Chemical class [Cl-].C=12C=CC(=[N+](CC)CC)C=C2OC2=CC(N(CC)CC)=CC=C2C=1C1=CC=CC=C1C(O)=O PYWVYCXTNDRMGF-UHFFFAOYSA-N 0.000 title description 5
- 125000003729 nucleotide group Chemical group 0.000 claims description 176
- 239000002773 nucleotide Substances 0.000 claims description 126
- 150000001875 compounds Chemical class 0.000 claims description 112
- 125000000217 alkyl group Chemical group 0.000 claims description 108
- 125000000547 substituted alkyl group Chemical group 0.000 claims description 89
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 51
- 238000012163 sequencing technique Methods 0.000 claims description 48
- 125000003545 alkoxy group Chemical group 0.000 claims description 42
- 229910052736 halogen Inorganic materials 0.000 claims description 41
- 150000002367 halogens Chemical class 0.000 claims description 41
- 125000005647 linker group Chemical group 0.000 claims description 40
- 125000003118 aryl group Chemical group 0.000 claims description 36
- 238000000034 method Methods 0.000 claims description 36
- 229910052799 carbon Inorganic materials 0.000 claims description 33
- 125000003107 substituted aryl group Chemical group 0.000 claims description 33
- 230000015572 biosynthetic process Effects 0.000 claims description 25
- -1 R 2 is H Chemical group 0.000 claims description 22
- 230000000903 blocking effect Effects 0.000 claims description 19
- 238000003786 synthesis reaction Methods 0.000 claims description 17
- 108091034117 Oligonucleotide Proteins 0.000 claims description 16
- SOWBFZRMHSNYGE-UHFFFAOYSA-N oxamic acid Chemical compound NC(=O)C(O)=O SOWBFZRMHSNYGE-UHFFFAOYSA-N 0.000 claims description 16
- 235000000346 sugar Nutrition 0.000 claims description 10
- 125000004356 hydroxy functional group Chemical group O* 0.000 claims description 9
- 238000004458 analytical method Methods 0.000 claims description 7
- CZPWVGJYEJSRLH-UHFFFAOYSA-N Pyrimidine Chemical compound C1=CN=CN=C1 CZPWVGJYEJSRLH-UHFFFAOYSA-N 0.000 claims description 6
- PYMYPHUHKUWMLA-LMVFSUKVSA-N Ribose Natural products OC[C@@H](O)[C@@H](O)[C@@H](O)C=O PYMYPHUHKUWMLA-LMVFSUKVSA-N 0.000 claims description 4
- HMFHBZSHGGEWLO-UHFFFAOYSA-N alpha-D-Furanose-Ribose Natural products OCC1OC(O)C(O)C1O HMFHBZSHGGEWLO-UHFFFAOYSA-N 0.000 claims description 4
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- HMFHBZSHGGEWLO-SOOFDHNKSA-N D-ribofuranose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H]1O HMFHBZSHGGEWLO-SOOFDHNKSA-N 0.000 claims description 2
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- 238000000926 separation method Methods 0.000 description 2
- 125000005017 substituted alkenyl group Chemical group 0.000 description 2
- 125000004426 substituted alkynyl group Chemical group 0.000 description 2
- HXJUTPCZVOIRIF-UHFFFAOYSA-N sulfolane Chemical compound O=S1(=O)CCCC1 HXJUTPCZVOIRIF-UHFFFAOYSA-N 0.000 description 2
- 229910052717 sulfur Inorganic materials 0.000 description 2
- 239000001117 sulphuric acid Substances 0.000 description 2
- 235000011149 sulphuric acid Nutrition 0.000 description 2
- RYYWUUFWQRZTIU-UHFFFAOYSA-K thiophosphate Chemical compound [O-]P([O-])([O-])=S RYYWUUFWQRZTIU-UHFFFAOYSA-K 0.000 description 2
- 229940113082 thymine Drugs 0.000 description 2
- 238000013519 translation Methods 0.000 description 2
- 229940035893 uracil Drugs 0.000 description 2
- WOMFWGRSYUKSSE-UHFFFAOYSA-N (1,3-dioxo-2-benzofuran-5-yl) acetate Chemical compound CC(=O)OC1=CC=C2C(=O)OC(=O)C2=C1 WOMFWGRSYUKSSE-UHFFFAOYSA-N 0.000 description 1
- DASBDJGFOOHLEB-UHFFFAOYSA-N 1,2,3,4,4a,5-hexahydroquinolin-7-ol Chemical compound C1CCNC2=CC(O)=CCC21 DASBDJGFOOHLEB-UHFFFAOYSA-N 0.000 description 1
- AACROBBFQRNFHE-UHFFFAOYSA-N 1,2,3,4-tetrahydroquinolin-5-ol Chemical class N1CCCC2=C1C=CC=C2O AACROBBFQRNFHE-UHFFFAOYSA-N 0.000 description 1
- IQQRAVYLUAZUGX-UHFFFAOYSA-N 1-butyl-3-methylimidazolium Chemical compound CCCCN1C=C[N+](C)=C1 IQQRAVYLUAZUGX-UHFFFAOYSA-N 0.000 description 1
- NJMWOUFKYKNWDW-UHFFFAOYSA-N 1-ethyl-3-methylimidazolium Chemical compound CCN1C=C[N+](C)=C1 NJMWOUFKYKNWDW-UHFFFAOYSA-N 0.000 description 1
- CDAWCLOXVUBKRW-UHFFFAOYSA-N 2-aminophenol Chemical compound NC1=CC=CC=C1O CDAWCLOXVUBKRW-UHFFFAOYSA-N 0.000 description 1
- CTGSQPRDMHCIMM-UHFFFAOYSA-N 3-(ethylamino)-4-methylphenol Chemical compound CCNC1=CC(O)=CC=C1C CTGSQPRDMHCIMM-UHFFFAOYSA-N 0.000 description 1
- MNUOZFHYBCRUOD-UHFFFAOYSA-N 3-hydroxyphthalic acid Chemical class OC(=O)C1=CC=CC(O)=C1C(O)=O MNUOZFHYBCRUOD-UHFFFAOYSA-N 0.000 description 1
- WJGUPVNQROJMKY-UHFFFAOYSA-N 4-[(1,3-dioxo-2-benzofuran-5-yl)oxy]butanoic acid Chemical compound OC(=O)CCCOC1=CC=C2C(=O)OC(=O)C2=C1 WJGUPVNQROJMKY-UHFFFAOYSA-N 0.000 description 1
- GJZWWIITSLTEET-UHFFFAOYSA-N C1=2C=C3C=C(C)C(NCC)=CC3=[O+]C=2C=C(NCC)C(C)=C1C1=CC(OCCCC(O)=O)=CC=C1C([O-])=O Chemical compound C1=2C=C3C=C(C)C(NCC)=CC3=[O+]C=2C=C(NCC)C(C)=C1C1=CC(OCCCC(O)=O)=CC=C1C([O-])=O GJZWWIITSLTEET-UHFFFAOYSA-N 0.000 description 1
- LNMRIJMCWQCPLP-UHFFFAOYSA-N C1=2C=C3C=C(C)C(NCC)=CC3=[O+]C=2C=C(NCC)C(C)=C1C1=CC=C(OCCCC(O)=O)C=C1C([O-])=O Chemical compound C1=2C=C3C=C(C)C(NCC)=CC3=[O+]C=2C=C(NCC)C(C)=C1C1=CC=C(OCCCC(O)=O)C=C1C([O-])=O LNMRIJMCWQCPLP-UHFFFAOYSA-N 0.000 description 1
- 230000006820 DNA synthesis Effects 0.000 description 1
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 1
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 1
- 208000005156 Dehydration Diseases 0.000 description 1
- OKKJLVBELUTLKV-MZCSYVLQSA-N Deuterated methanol Chemical compound [2H]OC([2H])([2H])[2H] OKKJLVBELUTLKV-MZCSYVLQSA-N 0.000 description 1
- BWGNESOTFCXPMA-UHFFFAOYSA-N Dihydrogen disulfide Chemical compound SS BWGNESOTFCXPMA-UHFFFAOYSA-N 0.000 description 1
- NYHBQMYGNKIUIF-UUOKFMHZSA-N Guanosine Chemical class C1=NC=2C(=O)NC(N)=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O NYHBQMYGNKIUIF-UUOKFMHZSA-N 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 238000007126 N-alkylation reaction Methods 0.000 description 1
- PXHVJJICTQNCMI-UHFFFAOYSA-N Nickel Chemical compound [Ni] PXHVJJICTQNCMI-UHFFFAOYSA-N 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- LGRFSURHDFAFJT-UHFFFAOYSA-N Phthalic anhydride Natural products C1=CC=C2C(=O)OC(=O)C2=C1 LGRFSURHDFAFJT-UHFFFAOYSA-N 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 239000007868 Raney catalyst Substances 0.000 description 1
- 229910000564 Raney nickel Inorganic materials 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- 239000005864 Sulphur Substances 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 125000002877 alkyl aryl group Chemical group 0.000 description 1
- 125000005600 alkyl phosphonate group Chemical group 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 150000001540 azides Chemical class 0.000 description 1
- 125000001797 benzyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])* 0.000 description 1
- 239000006177 biological buffer Substances 0.000 description 1
- 238000006664 bond formation reaction Methods 0.000 description 1
- JHIWVOJDXOSYLW-UHFFFAOYSA-N butyl 2,2-difluorocyclopropane-1-carboxylate Chemical compound CCCCOC(=O)C1CC1(F)F JHIWVOJDXOSYLW-UHFFFAOYSA-N 0.000 description 1
- 150000001732 carboxylic acid derivatives Chemical class 0.000 description 1
- 238000009903 catalytic hydrogenation reaction Methods 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 238000007385 chemical modification Methods 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 230000021615 conjugation Effects 0.000 description 1
- 239000013058 crude material Substances 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 125000004093 cyano group Chemical group *C#N 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 238000010511 deprotection reaction Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- WGLUMOCWFMKWIL-UHFFFAOYSA-N dichloromethane;methanol Chemical compound OC.ClCCl WGLUMOCWFMKWIL-UHFFFAOYSA-N 0.000 description 1
- 125000004177 diethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- 239000013024 dilution buffer Substances 0.000 description 1
- NAGJZTKCGNOGPW-UHFFFAOYSA-K dioxido-sulfanylidene-sulfido-$l^{5}-phosphane Chemical compound [O-]P([O-])([S-])=S NAGJZTKCGNOGPW-UHFFFAOYSA-K 0.000 description 1
- 239000001177 diphosphate Substances 0.000 description 1
- XPPKVPWEQAFLFU-UHFFFAOYSA-J diphosphate(4-) Chemical compound [O-]P([O-])(=O)OP([O-])([O-])=O XPPKVPWEQAFLFU-UHFFFAOYSA-J 0.000 description 1
- 235000011180 diphosphates Nutrition 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 238000000295 emission spectrum Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 125000003700 epoxy group Chemical group 0.000 description 1
- 238000001917 fluorescence detection Methods 0.000 description 1
- 125000005843 halogen group Chemical group 0.000 description 1
- BHEPBYXIRTUNPN-UHFFFAOYSA-N hydridophosphorus(.) (triplet) Chemical compound [PH] BHEPBYXIRTUNPN-UHFFFAOYSA-N 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 238000003018 immunoassay Methods 0.000 description 1
- 238000010324 immunological assay Methods 0.000 description 1
- 150000007529 inorganic bases Chemical class 0.000 description 1
- 229910010272 inorganic material Inorganic materials 0.000 description 1
- 239000011147 inorganic material Substances 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 229920000592 inorganic polymer Polymers 0.000 description 1
- 229910003480 inorganic solid Inorganic materials 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 239000000543 intermediate Substances 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- 238000011901 isothermal amplification Methods 0.000 description 1
- 239000011968 lewis acid catalyst Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 239000011859 microparticle Substances 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 238000001823 molecular biology technique Methods 0.000 description 1
- 239000003068 molecular probe Substances 0.000 description 1
- 125000000896 monocarboxylic acid group Chemical group 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 150000004712 monophosphates Chemical class 0.000 description 1
- 239000012038 nucleophile Substances 0.000 description 1
- 230000005257 nucleotidylation Effects 0.000 description 1
- 210000004940 nucleus Anatomy 0.000 description 1
- 238000002515 oligonucleotide synthesis Methods 0.000 description 1
- 150000007530 organic bases Chemical class 0.000 description 1
- 150000002894 organic compounds Chemical class 0.000 description 1
- 229920000620 organic polymer Polymers 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 150000002972 pentoses Chemical class 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N phenol group Chemical group C1(=CC=CC=C1)O ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 150000003003 phosphines Chemical class 0.000 description 1
- PTMHPRAIXMAOOB-UHFFFAOYSA-L phosphoramidate Chemical compound NP([O-])([O-])=O PTMHPRAIXMAOOB-UHFFFAOYSA-L 0.000 description 1
- 150000003016 phosphoric acids Chemical class 0.000 description 1
- XNGIFLGASWRNHJ-UHFFFAOYSA-L phthalate(2-) Chemical compound [O-]C(=O)C1=CC=CC=C1C([O-])=O XNGIFLGASWRNHJ-UHFFFAOYSA-L 0.000 description 1
- 150000003022 phthalic acids Chemical class 0.000 description 1
- 229920000867 polyelectrolyte Polymers 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 238000003752 polymerase chain reaction Methods 0.000 description 1
- 102000054765 polymorphisms of proteins Human genes 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 229910000027 potassium carbonate Inorganic materials 0.000 description 1
- LPNYRYFBWFDTMA-UHFFFAOYSA-N potassium tert-butoxide Chemical compound [K+].CC(C)(C)[O-] LPNYRYFBWFDTMA-UHFFFAOYSA-N 0.000 description 1
- CAKUHHOTEWNCDC-UHFFFAOYSA-M potassium;3,4-bis(ethoxycarbonyl)phenolate Chemical compound [K+].CCOC(=O)C1=CC=C([O-])C=C1C(=O)OCC CAKUHHOTEWNCDC-UHFFFAOYSA-M 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 238000002953 preparative HPLC Methods 0.000 description 1
- 150000003141 primary amines Chemical class 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 125000002572 propoxy group Chemical group [*]OC([H])([H])C(C([H])([H])[H])([H])[H] 0.000 description 1
- 125000001436 propyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 150000003230 pyrimidines Chemical class 0.000 description 1
- 238000010791 quenching Methods 0.000 description 1
- 239000012048 reactive intermediate Substances 0.000 description 1
- 238000011897 real-time detection Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 150000003839 salts Chemical group 0.000 description 1
- 238000007480 sanger sequencing Methods 0.000 description 1
- 238000007127 saponification reaction Methods 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 238000011896 sensitive detection Methods 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 229960001866 silicon dioxide Drugs 0.000 description 1
- AKEJUJNQAAGONA-UHFFFAOYSA-N sulfur trioxide Chemical compound O=S(=O)=O AKEJUJNQAAGONA-UHFFFAOYSA-N 0.000 description 1
- 150000005621 tetraalkylammonium salts Chemical class 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 125000002264 triphosphate group Chemical class [H]OP(=O)(O[H])OP(=O)(O[H])OP(=O)(O[H])O* 0.000 description 1
- UNXRWKVEANCORM-UHFFFAOYSA-N triphosphoric acid Chemical compound OP(O)(=O)OP(O)(=O)OP(O)(O)=O UNXRWKVEANCORM-UHFFFAOYSA-N 0.000 description 1
- 238000012070 whole genome sequencing analysis Methods 0.000 description 1
- 238000010626 work up procedure Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/78—Ring systems having three or more relevant rings
- C07D311/80—Dibenzopyrans; Hydrogenated dibenzopyrans
- C07D311/82—Xanthenes
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
- C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
- C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
- C07H19/06—Pyrimidine radicals
- C07H19/10—Pyrimidine radicals with the saccharide radical esterified by phosphoric or polyphosphoric acids
-
- 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
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
- C09B11/00—Diaryl- or thriarylmethane dyes
- C09B11/04—Diaryl- or thriarylmethane dyes derived from triarylmethanes, i.e. central C-atom is substituted by amino, cyano, alkyl
- C09B11/10—Amino derivatives of triarylmethanes
- C09B11/24—Phthaleins containing amino groups ; Phthalanes; Fluoranes; Phthalides; Rhodamine dyes; Phthaleins having heterocyclic aryl rings; Lactone or lactame forms of triarylmethane dyes
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
- G01N33/582—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with fluorescent label
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Biomedical Technology (AREA)
- Genetics & Genomics (AREA)
- Immunology (AREA)
- Physics & Mathematics (AREA)
- Microbiology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Cell Biology (AREA)
- Analytical Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Pyrane Compounds (AREA)
- Plural Heterocyclic Compounds (AREA)
- Saccharide Compounds (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Claims (15)
1. Forbindelse med formlen (I) og mesomerer deraf: hvor M+/" er en almindelig modion,
k er et heltal på fra 0 til 6, q er et heltal på fra 1 til 6, Ri er H eller alkyl, aryl eller en substitueret alkyl- eller substitueret arylgruppe, R2 er H, alkyl eller en substitueret alkylgruppe, halogen, carboxy, carboxamid, en hydroxy-eller alkoxygruppe, eller R2 sammen med Ri eller R5 er en carbon- eller heterosubstitueret kæde, der danner en ring, R3 er H, alkyl eller en substitueret alkylgruppe, halogen, carboxy, carboxamid, en hydroxy-eller alkoxygruppe eller R3 sammen med R4 eller R6 er en carbon- eller heterosubstitueret kæde, der danner en ring, R4 er H eller et alkyl, aryl eller substitueret alkyl- eller substitueret arylgruppe, R5 og R6 er H, alkyl eller substitueret alkylgruppe, halogen, hydroxy- eller alkoxygruppe, Rs er H, halogen, en hydroxy- eller alkoxygruppe, alkyl- eller substitueret alkylgruppe eller sammen med Ri er en carbon- eller heterosubstitueret carbonkæde, der danner en ring, R9 er H, halogen, en hydroxy- eller alkoxygruppe, en alkyl- eller substitueret alkylgruppe eller sammen med R4 er en carbon- eller heterosubstitueret carbonkæde, der danner en ring, R7 er OR11 eller NR11R12, hvor Ru og R12 uafhængigt er H, alkyl eller et substitueret alkyl, R13 er OR14 eller NR14R15, hvor R14 og R15 uafhængigt er H, alkyl eller et substitueret alkyl; aryl eller et substitueret aryl, og Ri6 og R17 uafhængigt er H eller alkyl, en aryl eller substitueret alkyl- eller substitueret arylgruppe.
2. Forbindelse ifølge krav 1, hvor Ri6 er alkyl og Rn er alkyl, eller Ri6 ogRn er H.
3. Forbindelse ifølge krav 1, hvor Ri, R4, R5, Ré, Rs og R9 alle er H, R2 og R3 er H eller CH3.
4. Forbindelse ifølge krav 1, hvor Ri er bundet til R2 eller R? via en kæde af CFb-grupper for at danne en ring, og R4 er bundet til R3 eller R9 via en kæde af CH2-grupper for at danne en ring.
5. Forbindelse ifølge krav 1, hvor én eller flere af Ri, R4, Rie og R17 er alkylgrupper substitueret med en S03'-gruppe.
6. Forbindelse ifølge krav 1-5, hvor R7 er OH.
7. Forbindelse ifølge krav 1-6, hvor q er 3.
8. Forbindelse ifølge krav 1-7, hvor R13 er OH eller NR14R15, hvor R14 er H eller alkyl og Ri5 er alkyl eller et substitueret alkyl.
9. Forbindelse ifølge krav 8, hvor forbindelsen er bundet til et nukleotid eller oligonukleotid via substitueret alkylgruppe R15.
10. Nukleotid eller oligonukleotid mærket med forbindelse ifølge krav 1-9.
11. Mærket nukleotid eller oligonukleotid ifølge krav 10, hvor mærket er bundet til C5-positionen af en pyrimidinbase eller C7-positionen af en 7-deazapurinbase via en linker-del.
12. Mærket nukleotid eller oligonukleotid ifølge krav 10 eller 11, der endvidere omfatter en 3'-OH-blokeringsgruppe kovalent bundet til nukleotidets ribose- eller deoxyribosesukker.
13. Kit, der omfatter to eller flere nukleotider, hvor mindst ét nukleotid er et mærket nukleotid ifølge krav 10-12.
14. Anvendelse af et nukleotid ifølge et hvilket som helst af kravene 10-12, et oligonukleotid ifølge krav 10-12 eller kit ifølge krav 13 i sekvensering, ekspressionsanalyse, hybridiseringsanalyse, genetisk analyse, RNA-analyse eller proteinbindings-assays.
15. Fremgangsmåde til syntese af forbindelse ifølge krav 1 startende fra phthalsyrederivat med formlen Xa eller Xb. hvor q er 1-6.
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PCT/EP2013/054793 WO2014135223A1 (en) | 2013-03-08 | 2013-03-08 | Rhodamine compounds and their use as fluorescent labels |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| DK2964624T3 true DK2964624T3 (da) | 2017-04-03 |
Family
ID=47882139
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| DK13709182.3T DK2964624T3 (da) | 2013-03-08 | 2013-03-08 | Rhodaminforbindelser og anvendelse heraf som fluorescerende mærker |
Country Status (7)
| Country | Link |
|---|---|
| EP (1) | EP2964624B1 (da) |
| CN (1) | CN105263918B (da) |
| AU (1) | AU2013380647B2 (da) |
| CA (1) | CA2902992C (da) |
| DK (1) | DK2964624T3 (da) |
| ES (1) | ES2620427T3 (da) |
| WO (1) | WO2014135223A1 (da) |
Families Citing this family (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB201414098D0 (en) | 2014-08-08 | 2014-09-24 | Illumina Cambridge Ltd | Modified nucleotide linkers |
| US10190162B2 (en) | 2014-10-23 | 2019-01-29 | Complete Genomics, Inc. | Signal confinement sequencing (SCS) and nucleotide analogues for signal confinement sequencing |
| CN107459482B (zh) * | 2016-06-03 | 2020-11-17 | 华东理工大学 | 一氧化氮供体、其制备及应用 |
| US10385214B2 (en) | 2016-09-30 | 2019-08-20 | Illumina Cambridge Limited | Fluorescent dyes and their uses as biomarkers |
| SG11201810611YA (en) * | 2016-12-22 | 2018-12-28 | Illumina Cambridge Ltd | Coumarin compounds and their uses as fluorescent labels |
| GB201711219D0 (en) | 2017-07-12 | 2017-08-23 | Illumina Cambridge Ltd | Short pendant arm linkers for nucleotides in sequencing applications |
| CN114671843B (zh) * | 2020-12-24 | 2023-07-04 | 郑州思昆生物工程有限公司 | 一种荧光化合物、荧光修饰核苷酸、试剂盒 |
| CN114671842B (zh) * | 2020-12-24 | 2024-05-28 | 郑州思昆生物工程有限公司 | 一种荧光化合物及其制备方法、荧光修饰核苷酸、试剂盒 |
| CN114656499B (zh) * | 2020-12-24 | 2026-04-21 | 郑州思昆生物工程有限公司 | 一种硅基罗丹明化合物及其制备方法、荧光修饰核苷酸、试剂盒 |
| US12359068B2 (en) * | 2023-02-10 | 2025-07-15 | Singular Genomics Systems, Inc. | Rhodamine fluorescent compounds and production methods thereof |
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| US6664061B2 (en) | 1999-06-25 | 2003-12-16 | Amersham Biosciences Ab | Use and evaluation of a [2+2] photoaddition in immobilization of oligonucleotides on a three-dimensional hydrogel matrix |
| AU2001279244A1 (en) | 2000-08-09 | 2002-02-18 | Motorola, Inc. | The use and evaluation of a (2+2) photocycloaddition in immobilization of oligonucleotides on a three-dimensional hydrogel matrix |
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| US8951938B2 (en) * | 2007-12-20 | 2015-02-10 | New York University | Combinatorial rosamine library to detect cell-state switching |
| WO2010033011A1 (en) * | 2008-09-19 | 2010-03-25 | Cancer Research Initiatives Foundation | Rosamine derivatives as agents for the treatment of cancer |
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| EP2964624B1 (en) | 2017-01-04 |
| AU2013380647A1 (en) | 2015-10-01 |
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