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PCR pre-packing reagent and reaction tube pre-loaded with PCR reagent

A pre-packaging and reaction tube technology, applied in the field of molecular biology, can solve the problems of complex PCR operation process, easy loss of activity, detection errors, etc., and achieve the effects of reducing storage and transportation costs, protecting activity, and reducing damage.

Inactive Publication Date: 2019-02-01
SUZHOU BAIYUAN GENT CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Therefore, the technical problem to be solved by the present invention is to overcome the defects that PCR reagents in the prior art are prone to lose activity during repeated freezing and thawing, and the PCR operation process is complex and prone to detection errors, thereby providing a PCR prepackaged Reagent

Method used

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  • PCR pre-packing reagent and reaction tube pre-loaded with PCR reagent
  • PCR pre-packing reagent and reaction tube pre-loaded with PCR reagent
  • PCR pre-packing reagent and reaction tube pre-loaded with PCR reagent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] This embodiment provides a PCR pre-packed reagent, which is obtained by freeze-drying the PCR reaction mixture and a lyoprotectant as raw materials. The preparation method of the PCR pre-packed reagent is as follows:

[0055] 1. Prepare PCR reaction mixture

[0056] Mix the PCR reaction reagents of 50×20 μl system to obtain 1000 μl of PCR reaction mixture, and the components of the PCR reaction mixture are shown in Table 1:

[0057] Table 1 Composition of PCR reaction mixture

[0058] Element

X 1

X 50

Final concentration

Taq enzyme (20U / μl)

1μl

50μl

1U / μl

Tris-HCl (300mM)

4μl

200μl

60mM

dNTPs (2mM)

2μl

100μl

0.2mM

MgCl 2 (120mM)

0.5μl

25μl

3mM

KCl (250mM)

4μl

200μl

50mM

(NH 4 ) 2 SO 4 (200mM)

0.5μl

25μl

5mM

SYBR Green I(40X)

0.5μl

25μl

1X

DEPC water

7.5μl

375μl

total

20μl

1000μl ...

Embodiment 2

[0071] This embodiment provides a PCR pre-packed reagent, which is obtained by freeze-drying the PCR reaction mixture and a lyoprotectant as raw materials. The preparation method of the PCR pre-packed reagent is as follows:

[0072] 1. Prepare PCR reaction mixture

[0073] Mix the PCR reaction reagents of the 50×20 μl system to obtain 1000 μl of the PCR reaction mixture, and the components of the PCR reaction mixture are shown in Table 2:

[0074] Table 2 Composition of PCR reaction mixture

[0075]

[0076]

[0077] 2. Weigh chitosan (purchased from sigma-aldrich, article number 9012-76-4) 0.15g, bovine serum albumin (purchased from sigma-aldrich, article number 9048-46-8) 0.10g, collagen (purchased from Shanghai Xinyu Biotechnology Co., Ltd., product number 9007-34-5) 0.05g, as a freeze-drying protection agent for PCR reaction solution.

[0078] 3. Add chitosan and bovine serum albumin to the 1000 μl PCR reaction mixture prepared in step 1, and mix evenly to obtain t...

Embodiment 3

[0089] This embodiment provides a PCR pre-packed reagent, which is obtained by freeze-drying the PCR reaction mixture and a lyoprotectant as raw materials. The preparation method of the PCR pre-packed reagent is as follows:

[0090] 1. Prepare PCR reaction mixture

[0091] Mix the PCR reaction reagents of 50×20 μl system to obtain 1000 μl of PCR reaction mixture, and the components in the PCR reaction mixture are shown in Table 3:

[0092] Table 3PCR reaction mixture composition

[0093]

[0094]

[0095] 2. Weigh chitosan oligosaccharide (purchased from Hubei Jusheng Technology Co., Ltd., article number 8411-57-8) 0.03g, chitosan (purchased from sigma-aldrich, article number 9012-76-4) 0.02g, bovine serum white 0.0125 g of protein (purchased from sigma-aldrich, product number 9048-46-8) was used as a lyoprotectant for the PCR reaction solution.

[0096]3. Add chitosan and bovine serum albumin to the 1000 μl PCR reaction mixture prepared in step 1, and mix evenly to ob...

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Abstract

The invention discloses a PCR pre-packing reagent. The PCR pre-packing reagent is a lyophilized reagent, the lyophilized reagent is composed of a PCR reaction mixed solution and a cryoprotectant, andthe weight of the cryoprotectant is 6.25-30% (w / v) of the volume of the PCR reaction mixed solution. The PCR pre-packing reagent simplifies the PCR operation step and improves the experimental efficiency; and the PCR pre-packing reagent also has a high amplification activity and a high quality stability, and can be stored at room temperature for a long term to reduce the storage and transportationcost of the reagent. The invention also provides a reaction tube pre-loaded with the PCR reagent, the reaction chamber of the reaction tube is filled with the PCR pre-packing reagent, an identification code for distinguishing different reaction tubes is arranged on the surface of the reaction tube, and the identification code does not block lights transmitting out of the reaction tube from the reaction chamber. Distinguishing of the different reaction tubes with the identification code is beneficial to achieve automatic matching of PCR detection results with sample and index information, so the automatic operation of PCR detection experiments is realized.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a PCR pre-packed reagent and a reaction tube for pre-packed PCR reagent. Background technique [0002] Polymerase chain reaction (PCR), also known as in vitro amplification technology, is a new technology for rapid amplification of specific DNA sequences in vitro, designed according to the characteristics of DNA replication in organisms. It was first developed in 1985 by Kary Mullis, Department of Human Genetics, Cetus Corporation, University of California, Los Angeles, and Howghes School of Medicine. This technology has changed the mode of traditional molecular cloning, that is, it does not amplify through the purification of the target gene and its in vitro recombination into in vitro DNA with autonomous replication ability. It can make several copies of the template sequence within a few hours, or even One DNA molecule amplifies 10 7 -10 9 times, greatly improving the rate ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/686C12M1/00C12M1/24
CPCC12Q1/686C12Q2527/125
Inventor 车团结徐红李琳陈游
Owner SUZHOU BAIYUAN GENT CO LTD
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