CA2856345C - Device for thermal convection polymerase chain reaction - Google Patents

Device for thermal convection polymerase chain reaction Download PDF

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Publication number
CA2856345C
CA2856345C CA2856345A CA2856345A CA2856345C CA 2856345 C CA2856345 C CA 2856345C CA 2856345 A CA2856345 A CA 2856345A CA 2856345 A CA2856345 A CA 2856345A CA 2856345 C CA2856345 C CA 2856345C
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Canada
Prior art keywords
test tube
diameter section
passage
heat dissipation
dissipation base
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Active
Application number
CA2856345A
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French (fr)
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CA2856345A1 (en
Inventor
Cheng SU
Ping Hua Teng
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Genereach Biotechnology Corp
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Genereach Biotechnology Corp
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Publication date
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Publication of CA2856345A1 publication Critical patent/CA2856345A1/en
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Classifications

    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L7/00Heating or cooling apparatus; Heat insulating devices
    • B01L7/52Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
    • B01L7/525Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples with physical movement of samples between temperature zones
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00Additional constructional details
    • B01L2300/08Geometry, shape and general structure
    • B01L2300/0832Geometry, shape and general structure cylindrical, tube shaped
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00Additional constructional details
    • B01L2300/08Geometry, shape and general structure
    • B01L2300/0832Geometry, shape and general structure cylindrical, tube shaped
    • B01L2300/0838Capillaries
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00Additional constructional details
    • B01L2300/18Means for temperature control
    • B01L2300/1805Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00Additional constructional details
    • B01L2300/18Means for temperature control
    • B01L2300/1861Means for temperature control using radiation
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00Moving or stopping fluids
    • B01L2400/04Moving fluids with specific forces or mechanical means
    • B01L2400/0403Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/0442Moving fluids with specific forces or mechanical means specific forces thermal energy, e.g. vaporisation, bubble jet
    • B01L2400/0445Natural or forced convection
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L9/00Supporting devices; Holding devices
    • B01L9/06Test-tube stands; Test-tube holders
    • B01L9/065Test-tube stands; Test-tube holders specially adapted for capillary tubes

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Molecular Biology (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

Provided in the present invention is a device for thermal convection polymerase chain reaction (PCR). The device is used for bearing a test tube such that PCR is performed in the test tube. The device comprises a heat radiating base which has a body and a passage provided through the body to be used for the insertion of the test tube, wherein the passage has a large diameter section and a small diameter section positioned below the large diameter section. Therefore, it can be ensured that during the PCR process the surface temperature of the mixed liquid is lower than the temperature required in a primer annealing reaction.

Description

DEVICE FOR THERMAL CONVECTION POLYMERASE CHAIN
REACTION
BACKGROUND OF THE INVENTION
1. Field of the Invention The present invention relates generally to devices for use in polymerase chain reaction (hereinafter referred to as "PCR") and more particularly, to a device for holding a test tube for insulated isothermal PCR.
2. Description of the Related Art In the field of biotechnology, polymerase chain reaction (PCR) is a well-known technology used to amplify specific nucleic acid sequences. The PCR

process comprises three major steps including denaturation, primer annealing and extension. which require different reaction temperatures. The required temperature for the denaturation step is typically in a range between 90 C and 97 C. The required temperature for the primer annealing step will depend on the melting temperature of the primer used. Typically, the annealing temperature ranges from 35 C to 65 C. The required temperature for the extension step is typically about 72 C.
The insulated isothermal PCR is based on Rayleigh-Benard convection, which is driven by buoyancy when heating fluid layer from below, is a common physical phenomenon. The insulated isothermal PCR is generally performed by immersing the bottom of a test tube which contains a mixed liquid into a hot water in such a way that the rest of the test tube is exposed to atmosphere at room temperature for heat dissipation. As a result, the temperature of the mixed liquid will gradually decrease from the bottom of the mixed liquid having a temperature of about 97 C
toward the surface of the mixed liquid having a temperature of about 35 C.

=
Because of the temperature gradient, the heat convection is induced, such that the mixed liquid will flow through various regions having different temperatures and then undergo different reaction steps.
In the conventional device for performing a convection PCR, because the portion of the test tube, which is exposed to the ambient air at room temperature for heat dissipation, has a low heat dissipating rate, the temperature at the surface of the mixed liquid will become higher and higher due to the increment of the heating time.
As a result, the temperature at the surface of the mixed liquid may rise to a degree higher than the required temperature suitable for conducting the primer annealing step before the convection PCR has been performed completely. Under this circumstance, the polymerase chain reaction may break, such that a desired, large amount of copies of specific nucleic acid sequences may not be obtained.
SUMMARY OF THE INVENTION
The present invention has been accomplished in view of the above-noted circumstances. It is the primary objective of the present invention to provide a device for insulated isothermal PCR, which can ensure that the temperature at the surface of the mixed liquid is lower than the temperature suitable for conducting the primer annealing step in the PCR process.
To achieve the above-mentioned objective, the device provided by the present invention is adapted for holding a test tube in which an insulated isothermal polymerase chain reaction is performed, which comprises a heat radiating base having a body provided with a passage for insertion of the test tube. The passage has a large diameter section and a small diameter section located below the large diameter section.
By means of the design of the present invention, the temperature at the surface of the mixed liquid can be kept in a degree lower than the temperature suitable for conducting the primer annealing step in the PCR process.
According to one aspect of the invention, it is provided that a device for holding a test tube in which insulated isotheimal polymerase chain reaction is performed, the = device comprising: a heat radiating base having a body provided with a passage for insertion of the test tube, wherein the passage has a large diameter section and a small diameter section located below the large diameter section; wherein the heat radiating base dissipates heat from the test tube which is inserted in the passage; the heat radiating base leaves a bottom of the test tube exposed outside to be heated.
According to another aspect of the present invention, there is provided a device for holding a test tube in which thelinal convection polymerase chain reaction is performed, and for dissipating heat from the test tube, the device comprising:

a heat dissipation base having a body provided with a passage for insertion of the test tube, wherein the passage has a large diameter section and a small diameter section located below the large diameter section;
wherein the test tube and the heat dissipation base are separated by an air space; the heat dissipation base dissipates heat from the test tube which is inserted in the passage;
the heat dissipation base leaves a bottom of the test tube exposed outside to be heated.
BRIEF DESCRIPTION OF THE DRAWINGS
The present invention will become more fully understood from the detailed description given herein below and the accompanying drawings which are given by way of illustration only, and thus are not limitative of the present invention, and wherein:
FIG. 1 is a schematic drawing showing a device for insulated isothermal PCR
according to a preferred embodiment of the present invention; and FIG. 2 is an exploded view of the device for insulated isothermal PCR of the preferred embodiment of the present invention.
3 DETAILED DESCRIPTION OF THE INVENTION
As shown in FIG. 1, a device 10 for insulated isothermal PCR mainly comprises a heat radiating base 20 and a tube rack 30. The heat radiating base 20 includes a body 22 provided with a passage 24 for insertion of a test tube 12. The passage 24 has a large diameter section 241 and a small diameter section 242 located below the large diameter section 241. Because a middle section 123 and an upper section 122 of the test tube 12 are located inside the passage 24 of the heat radiating base 20 and the heat radiating base 20 is made of a material having a high heat transfer coefficient, such as aluminum, the heat energy of the mixed liquid of PCR will be transferred through the ambient atmosphere to the heat radiating base 20 for heat dissipation efficiently, such that during the PCR process the temperature at the surface of the mixed liquid can be maintained at a degree of about 10-55 C, which is lower 3a than the temperature suitable for performing the primer annealing step, thereby preventing the PCR from being interrupted due to high temperature at the mixed liquid level. In addition, the heat dissipation of the mixed liquid at the region corresponding to the large diameter section 241 will be lower than that at the region corresponding to the small diameter section 242. It is revealed by experiments that the configuration of the heat radiating base 20 provided by the present invention has a heat-dissipating effect helpful for conducting the insulated isothermal PCR.
In order to stably mount the test tube 12 in the heat radiating base 20, the tube rack 30 can be further provided on the heat radiating base 20. The tube rack 30 is provided with a receiving hole 32 for insertion of the test tube 12 to stably position the test tube 12, thereby preventing the outer wall surface of the test tube 12 from contacting the wall surface of the passage 24 of the heat radiating base 20 so as to avoid that the temperature of the mixed liquid drops too quickly.
In practice, the receiving hole 32 of the tube rack 30 may be configured, in succession order from a top thereof toward a bottom thereof, a large diameter section 321, a shoulder 322 and a small diameter section 323. in which the shoulder 322 is adapted for stopping a shoulder 121 of the test tube 12 such that the test tube 12 can be stably positioned. In addition, a support seat 40 is provided below the heat radiating base 20. The bottom 124 of the test tube 12 is heated by a heat source (not shown in the drawings) to keep the temperature of the mixed liquid of PCR inside the bottom 124 in a range about 90 C to 97 C.
Further, the bottom end 301 of the tube rack 30 is inserted into the passage 24 of the heat radiating base 20, such that the tube rack 30 is stably mounted on the heat radiating base 20.
Furthermore, in the preferred embodiment of the present invention, the
4 diameter of the small diameter section 242 of the passage 24 is configured to gradually and downwardly decrease. According to many experiments and modifications, it is found that using this configuration to dissipate heat can yield a highest reaction efficiency of PCR. The aforesaid experiments for PCR were conducted in seven different environmental temperatures ranging from 10 C to 40 C with a condition that the mixed liquid inside the bottom 124 of the test tube 12 was heated to 93 C
to 97 C.
The temperature of the heat radiating base 20 measured ranges from 36 C to 53 C, and the temperature at the mixed liquid level measured ranges from 36 C to 53 C;
therefore, the PCR is performed smoothly.
The invention being thus described, it will be obvious that the same may be varied in many ways. For example, as shown in FIG. 2, the heat radiating base 20 can be provided with a plurality of passages 24 for holding a plurality of test tubes 12 for simultaneously performing polymerase chain reactions. Such variations are not to be regarded as a departure from the spirit and scope of the invention, and all such modifications as would be obvious to one skilled in the art are intended to be included within the scope of the following claims.
5

Claims (6)

Claims
1. A device for holding a test tube in which thermal convection polymerase chain reaction is performed, and for dissipating heat from the test tube, the device comprising:
a heat dissipation base having a body provided with a passage for insertion of the test tube, wherein the passage has a large diameter section and a small diameter section located below the large diameter section;
wherein the test tube and the heat dissipation base are separated by an air space;
the heat dissipation base dissipates heat from the test tube which is inserted in the passage; the heat dissipation base leaves a bottom of the test tube exposed outside to be heated.
2. The device of claim 1, further comprising a tube rack mounted on the heat dissipation base and provided with a receiving hole for insertion of the test tube.
3. The device of claim 2, wherein the receiving hole of the tube rack includes, in successive order from a top of the receiving hole toward a bottom of the receiving hole, a large diameter section, a shoulder, and a small diameter section, wherein the shoulder of the receiving hole is adapted to stop a shoulder of the test tube.
4. The device of claim 2, wherein the tube rack has a bottom end inserted into the passage of the heat dissipation base.
5. The device of any one of claims 1 to 4, wherein the small diameter section of the passage of the heat dissipation base has a gradually decreasing diameter from the top to the bottom of said passage of said heat dissipation base.
6. The device of claim 1, wherein the heat dissipation base is made of metal.
CA2856345A 2011-11-22 2011-11-22 Device for thermal convection polymerase chain reaction Active CA2856345C (en)

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/CN2011/001941 WO2013075263A1 (en) 2011-11-22 2011-11-22 Device for thermal convection polymerase chain reaction

Publications (2)

Publication Number Publication Date
CA2856345A1 CA2856345A1 (en) 2013-05-30
CA2856345C true CA2856345C (en) 2017-10-24

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EP (1) EP2784150B1 (en)
KR (1) KR101691466B1 (en)
CN (1) CN103649301B (en)
CA (1) CA2856345C (en)
IN (1) IN2014CN03868A (en)
WO (1) WO2013075263A1 (en)

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10487301B2 (en) * 2015-05-12 2019-11-26 Xiamen University Reaction tube for nucleic acid amplification capable of controlling liquid circulation path
CN105670924A (en) * 2016-03-01 2016-06-15 上海理工大学 Natural convection PCR (Polymerase Chain Reaction) microsystem
CN107964507B (en) 2016-10-18 2022-09-27 财团法人工业技术研究院 Thermal convection polymerase chain reaction device and optical detection method thereof
CN108410688A (en) 2017-02-09 2018-08-17 克雷多生物医学私人有限公司 A kind of device of heat convection type PCR
CN108949545A (en) * 2018-08-16 2018-12-07 上海海洋大学 A kind of novel nucleic acids isothermal amplification component
CN109294901A (en) * 2018-11-01 2019-02-01 福建省博凯科技有限公司 One-part form thermal convection PCR instrument and control method
CN109401948A (en) * 2018-11-01 2019-03-01 福建省博凯科技有限公司 One-part form thermal convection PCR instrument
CN109554295B (en) * 2019-01-21 2022-03-29 武汉理工大学 PCR amplification and disease detection device for ocean-going crew

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CA2130013C (en) 1993-09-10 1999-03-30 Rolf Moser Apparatus for automatic performance of temperature cycles
KR100488281B1 (en) * 2001-09-15 2005-05-10 아람 바이오시스템 주식회사 Method and apparatus for amplification of nucleic acid sequences by using thermal convection
DE102004039092B4 (en) 2004-08-11 2008-04-24 Deutsches Zentrum für Luft- und Raumfahrt e.V. Locking device for linear actuators
US8153064B2 (en) * 2007-03-22 2012-04-10 Doebler Ii Robert W Systems and devices for isothermal biochemical reactions and/or analysis
GB0715170D0 (en) 2007-08-03 2007-09-12 Enigma Diagnostics Ltd Reaction vessel
US20100124779A1 (en) * 2008-11-18 2010-05-20 Roche Diagnostics Operations, Inc. Two-Step Moulded Capillary
JP5865261B2 (en) * 2010-01-12 2016-02-17 アーラム バイオシステムズ インコーポレイテッド Two-stage thermal convection device and method of use

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Publication number Publication date
CA2856345A1 (en) 2013-05-30
EP2784150A4 (en) 2015-08-12
KR20140040262A (en) 2014-04-02
EP2784150B1 (en) 2019-05-15
CN103649301B (en) 2015-03-11
IN2014CN03868A (en) 2015-10-16
CN103649301A (en) 2014-03-19
KR101691466B1 (en) 2016-12-30
WO2013075263A1 (en) 2013-05-30
EP2784150A1 (en) 2014-10-01

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Effective date: 20140520