WO2017106917A2 - Récipient de réaction et ensemble couvercle - Google Patents

Récipient de réaction et ensemble couvercle Download PDF

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Publication number
WO2017106917A2
WO2017106917A2 PCT/AU2016/051268 AU2016051268W WO2017106917A2 WO 2017106917 A2 WO2017106917 A2 WO 2017106917A2 AU 2016051268 W AU2016051268 W AU 2016051268W WO 2017106917 A2 WO2017106917 A2 WO 2017106917A2
Authority
WO
WIPO (PCT)
Prior art keywords
cap
reaction container
barrier fluid
reagent
nucleic acid
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.)
Ceased
Application number
PCT/AU2016/051268
Other languages
English (en)
Other versions
WO2017106917A3 (fr
Inventor
John Corbett
SNR John CORBETT
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.)
BIO MOLECULAR SYSTEMS Pty Ltd
Original Assignee
BIO MOLECULAR SYSTEMS Pty Ltd
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
Priority claimed from AU2015905312A external-priority patent/AU2015905312A0/en
Application filed by BIO MOLECULAR SYSTEMS Pty Ltd filed Critical BIO MOLECULAR SYSTEMS Pty Ltd
Publication of WO2017106917A2 publication Critical patent/WO2017106917A2/fr
Publication of WO2017106917A3 publication Critical patent/WO2017106917A3/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5021—Test tubes specially adapted for centrifugation purposes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/06—Fluid handling related problems
    • B01L2200/0673—Handling of plugs of fluid surrounded by immiscible fluid
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/06—Fluid handling related problems
    • B01L2200/0689—Sealing
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/04—Closures and closing means
    • B01L2300/041—Connecting closures to device or container
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/04—Closures and closing means
    • B01L2300/046—Function or devices integrated in the closure
    • B01L2300/047—Additional chamber, reservoir
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/0681—Filter
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/508—Rigid containers without fluid transport within
    • B01L3/5082—Test tubes per se
    • B01L3/50825—Closing or opening means, corks, bungs

Definitions

  • the barrier fluid comprises a specific gravity between 0.5 and 0.95 over a temperature range of -20 to 100 e C.
  • the barrier fluid may be a silicone wax.
  • the wax has a molecular weight of about 12,600 amu.
  • the wax has a melting point in the range of about 40 to 50 e C, preferably 40 to 45 e C, more preferably about 43 e C.
  • the silicone wax is an active organo silicone wax copolymer.
  • a suitable wax may be sourced from Genesee Polymers Corporation (GP-58). The preferred silicone wax of the invention, when in a molten form, has the viscosity, specific gravity, and/or surface tension as described above.
  • a plurality of reaction containers are joined together in a circular array.
  • the array can be configured to hold fewer or more than 20 of the reaction containers as required, such as 30, 40, 50, 60, 70, 80, 90, or 100 reaction containers.
  • the reaction containers are angled at 80 degrees from the horizontal.
  • the circular array can be configured with any number of reaction containers as required and to suit the desired application.
  • the circular array of reaction containers is particularly suited to be loaded into the thermocycler apparatus of PCT Publication WO 2015/054733 (incorporated herein by reference).
  • the reaction container and cap assembly as described herein is particularly suited for automated loading in the thermocycler apparatus of WO 2015/054733, whereby the container and cap assembly can be placed in the thermocycler and loaded by a robot without further manipulation by a user.
  • a target nucleic acid can be provided within the reaction container, the target nucleic acid being separated from the atmosphere, the pre-loaded reagent, and the nucleic acid amplification reagents by use of 3 or more barrier fluid seals.
  • the barrier fluid of each individual layer of a plurality of seals can be the same or different, and can be chosen such that the seal permits transport of a fluid from one side to the other only when a predetermined centrifugal force is applied, and/or only when a predetermined temperature is reached during a heating cycle. Combinations of the above will be apparent to the skilled person.
  • the reaction container is suitable for use in a thermocycler system and method thereof, for example as described in WO 2015/054733, the contents of which are incorporated herein by reference.
  • the thermocycler comprises a rotatable platform formed, at least in part, of a material which is heatable in response to being exposed to electromagnetic energy; and an electromagnetic energy source adapted to direct
  • the present invention provides a thermocycler system, the system comprising: a) a rotatable platform containing at least one reaction container according to the second aspect, wherein the rotatable platform is formed, at least in part, of a material which is heatable in response to being exposed to electromagnetic energy; and b) an electromagnetic energy source adapted to direct electromagnetic energy at the rotatable platform, wherein the electromagnetic energy source surrounds a sufficient amount of the rotatable platform in order to heat the entire platform substantially simultaneously.
  • the present invention provides a method of cycling one or more reaction containers according to the second aspect between predetermined
  • At least some of the internal surfaces of the upper portion of the cap (and/or the upper portion of the reaction container) have a surface covering of nucleic acid amplification reagents in a 'dehydrated' or powdered form. In some embodiments, at least some of the internal surfaces of the upper portion of the cap (and/or the upper portion of the reaction container) also have a surface covering of the pre-loaded reagent in a 'dehydrated' or powdered form. In some embodiments, the 'dehydrated' or powdered particles are adhered to the internal surface of the upper portion of the cap (and/or the upper portion of the reaction container) by electrostatic forces. In other embodiments, the 'dehydrated' or powdered particles are adhered to the internal surface of the upper portion of the cap (and/or the upper portion of the reaction container) by van der Waals forces.
  • the first pre-loaded reagent and/or second pre-loaded reagent are selected independently from the group consisting of: an enzyme with polymerase activity (such as a DNA polymerase, Taq polymerase or reverse transcriptase), an enzyme, a nucleic acid, a nucleic acid primer, a buffer, a salt, a deoxyribonucleotide, a biological material, a preservative, an initiator, an acid, a nucleotide, a nucleoside triphosphate, or combinations thereof.
  • an enzyme with polymerase activity such as a DNA polymerase, Taq polymerase or reverse transcriptase
  • an enzyme such as a DNA polymerase, Taq polymerase or reverse transcriptase
  • an enzyme such as a DNA polymerase, Taq polymerase or reverse transcriptase
  • an enzyme such as a DNA polymerase, Taq polymerase or reverse transcriptase
  • an enzyme such as a DNA polymerase, Taq polymerase
  • the second barrier fluid is a silicon oil.
  • the lower portion contains a third pre-loaded reagent and/or a third barrier fluid positioned above said second barrier fluid.
  • the cap of the invention, the reaction container of the invention, or the assembly of the container and cap is particularly suited for use in conducting nucleic acid amplification reactions such as the polymerase chain reaction (PCR) and the ligase chain reaction (LCR).
  • PCR polymerase chain reaction
  • LCR ligase chain reaction
  • the cap/container/assembly of the invention can be used in a wide variety of systems which require multiple or cyclic chemical reactions to produce a desired product.
  • the sample to be analysed or reacted is a nucleic acid such as DNA or RNA containing sample.
  • PCR reagents 5 e.g., dNTPs, MgCI 2 , primers, probes, reaction buffers and sample
  • These reagents 5 sit on top of the silicon oil layer 4 and are physically separated from the Taq DNA polymerase 3 that sits at the bottom of the container 1 ( Figure 1 C), which in the case of Figures 1 B and 1 C is shown as a droplet, but could instead be a layer at the base of the reaction container 1 .
  • the reaction container 1 is heated to 60 e C and centrifuged at a speed of greater than about 1 ,000 rpm. This has the effect of mixing all the aqueous volumes together at 60 e C, which is equivalent to a "Hot start" qPCR reaction ( Figure 1 D).
  • Figure 2 shows the reaction container 1 sealed with a closure 6.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Hematology (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Devices For Use In Laboratory Experiments (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

La présente invention concerne un récipient de réaction qui est approprié pour mener une PCR. En particulier, l'invention concerne un récipient de réaction préchargé comprenant un réactif qui est recouvert d'un fluide-barrière visqueux fluide, le fluide-barrière visqueux enfermant hermétiquement le réactif vis-à-vis de l'atmosphère. Un ou plusieurs réactifs de réaction sont ajoutés au récipient de réaction préchargé contenant et le récipient est centrifugé pour combiner le ou les réactifs de réaction et le réactif préchargé. Le réactif préchargé est stable à des températures entre 10 et 40 °C pendant de nombreux mois lorsqu'il est plongé dans le fluide-barrière ou est étanche à l'atmosphère par le fluide-barrière. La présente invention concerne également un couvercle en vue d'une mise en prise coopérative avec un récipient de réaction, le couvercle comprenant une partie supérieure pour recevoir un échantillon et une partie inférieure conçue pour entrer en contact hermétique avec l'ouverture d'entrée du récipient de réaction afin de définir une chambre de réaction entre le couvercle et le récipient de réaction. La partie supérieure et la partie inférieure du couvercle sont en communication fluidique par un conduit conçu pour retenir sensiblement au moins un fluide-barrière dans la chambre de réaction.
PCT/AU2016/051268 2015-12-22 2016-12-22 Récipient de réaction et ensemble couvercle Ceased WO2017106917A2 (fr)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
AU2015905312A AU2015905312A0 (en) 2015-12-22 Reaction container
AU2015905312 2015-12-22
AU2016901589 2016-04-29
AU2016901589A AU2016901589A0 (en) 2016-04-29 Reaction container and cap assembly

Publications (2)

Publication Number Publication Date
WO2017106917A2 true WO2017106917A2 (fr) 2017-06-29
WO2017106917A3 WO2017106917A3 (fr) 2017-07-27

Family

ID=59088670

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/AU2016/051268 Ceased WO2017106917A2 (fr) 2015-12-22 2016-12-22 Récipient de réaction et ensemble couvercle

Country Status (1)

Country Link
WO (1) WO2017106917A2 (fr)

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2018174908A (ja) * 2017-08-24 2018-11-15 ヤマトエスロン株式会社 試薬カセット
JP2019118302A (ja) * 2017-12-29 2019-07-22 ヤマトエスロン株式会社 Ｐｃｒ用容器及び試薬入りｐｃｒ用容器
CN111420622A (zh) * 2020-04-30 2020-07-17 南通合瑞源生物科技有限公司 一种用于核酸合成纯化高通量多孔过滤板
WO2022219164A1 (fr) * 2021-04-14 2022-10-20 Synhelix Unité de réaction tubulaire
EP4647167A1 (fr) * 2024-05-08 2025-11-12 Relco Solutions Limited Récipient de réaction

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4417981A (en) * 1981-05-04 1983-11-29 Becton, Dickinson And Company Blood phase separator device
US5780248A (en) * 1993-07-15 1998-07-14 Ortho Diagnostic Systems, Inc. Foil sealed cassette for agglutination reactions and liner therefor
US6428962B1 (en) * 2001-02-12 2002-08-06 Dna Analysis, Inc. Nucleic acid collection barrier method and apparatus
WO2012016720A1 (fr) * 2010-08-05 2012-02-09 Vibod Gmbh Colonnes inédites utilisables en vue de l'incubation et de l'isolement d'échantillons chimiques et/ou biologiques

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2018174908A (ja) * 2017-08-24 2018-11-15 ヤマトエスロン株式会社 試薬カセット
JP2019118302A (ja) * 2017-12-29 2019-07-22 ヤマトエスロン株式会社 Ｐｃｒ用容器及び試薬入りｐｃｒ用容器
CN111420622A (zh) * 2020-04-30 2020-07-17 南通合瑞源生物科技有限公司 一种用于核酸合成纯化高通量多孔过滤板
WO2022219164A1 (fr) * 2021-04-14 2022-10-20 Synhelix Unité de réaction tubulaire
EP4647167A1 (fr) * 2024-05-08 2025-11-12 Relco Solutions Limited Récipient de réaction

Also Published As

Publication number Publication date
WO2017106917A3 (fr) 2017-07-27

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