PCR (polymerase chain reaction) reaction solution and kit, and PCR method

A reaction solution and solution technology, applied in the biological field, can solve problems such as affecting use and reagent contamination, and achieve the effects of reducing configuration time, improving amplification efficiency, and saving detection time

Inactive Publication Date: 2015-02-04
FUJIAN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In conventional PCR amplification system configuration, DNTP, Buffer, rTaq enzyme and other reagents are often stored separately. When PCR amplification is to be performed, the samples are added to the same

Method used

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  • PCR (polymerase chain reaction) reaction solution and kit, and PCR method
  • PCR (polymerase chain reaction) reaction solution and kit, and PCR method
  • PCR (polymerase chain reaction) reaction solution and kit, and PCR method

Examples

Experimental program
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Effect test

Embodiment 1

[0016] A PCR reaction solution, prepared from the following raw materials: 6-8 μl dimethyl sulfoxide, 6-8 μl bovine serum albumin solution with a mass volume percentage of 0.1%, 9-11 μl 10mM DNTP solution, 18-22 μl 10×PCR buffer, 2-4 μl bromophenol blue solution, 45-55 μl solute volume percent 30% glycerol solution and 4-6 μl rTaq enzyme.

[0017] In the traditional preparation of PCR reaction solution, DNTP, Buffer, rTaq enzyme and other reagents are mixed to prepare a PCR mixture that can be used directly. If the PCR mixture is stored at -20°C, the activity of rTaq enzyme will decrease. Therefore, the present invention adds a certain proportion of glycerin solution and the combination of BSA and DMSO to the mixed solution to ensure the stability and activity of Tag enzyme and reduce the formation of primer dimers.

[0018] A PCR reaction solution, the reaction solution is mixed by the following components: 6 μl dimethyl sulfoxide, 6 μl bovine serum albumin solution with a ma...

Embodiment 2

[0021] A PCR reaction solution, the reaction solution comprising 7.5 μl dimethyl sulfoxide, 7.5 μl mass volume percentage of 0.1% bovine serum albumin solution, 10 μl 10mM DNTP solution, 20 μl 10×PCR buffer, 5 μl bromophenol blue solution, 47 μl solute volume Percentages are 30% glycerol solution and 5 μl rTaq enzyme. The prepared solution was mixed and centrifuged, and stored in a -20°C refrigerator.

[0022] Further, in the above PCR reaction solution, the 10×PCR buffer contains 50-150mM Tris-Hcl with a pH of 8.3, 250-750mM Kcl and 5-20mM Mgcl 2 .

[0023] Further, in the above PCR reaction solution, the bromophenol blue solution contains 30% by volume of glycerin and 0.05% by mass volume of bromophenol blue powder.

[0024] A PCR kit, characterized in that it comprises any one of the above PCR reaction solutions.

[0025] Use of the above PCR reaction solution: Take out the 2×PCR reaction solution from the refrigerator, and configure the PCR system. The configuration of ...

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Abstract

The invention relates to the technical field of biology, particularly a PCR (polymerase chain reaction) reaction solution and kit, and a PCR method. The PCR reaction solution is prepared from dimethyl sulfoxide (DMSO), bovine serum albumin (BSA), dNTP (deoxyribonucleotide triphosphate), 10*PCR buffer, bromphenol blue solution, glycerol solution with the solute volume percent of 30%, and rTaq enzyme. The PCR reaction solution for PCR experiments is optimized; especially a certain proportion of glycerol solution is added, and the combination of the BSA and DMSO is added to ensure the stability and activity of the Tag enzyme, reduce the formation of the primer dimer and shorten the PCR reaction solution configuration time; and in the PCR product detection process by agarose gel electrophoresis, the detection can be performed only by adding the nucleic acid dye, thereby saving abundant time for detecting the PCR product and enhancing the PCR amplification efficiency.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a PCR reaction solution, a kit and a PCR method. Background technique [0002] Polymerase chain reaction (PCR) is a technique for rapidly amplifying specific DNA fragments in vitro, and it is one of the most important means of modern molecular biology. Its basic principle is based on the mechanism of somatic cell replication established by the DNA double helix model won the Nobel Prize in Physiology in 1953 and the characteristic that double-stranded DNA can undergo denaturation and renaturation in vitro with temperature changes. In 1985, Randll.K.Skaii established the polymerase chain reaction technology using Klenow DNA polymerase and applied it to the genetic diagnosis of sickle cell anemia. It was a revolutionary breakthrough in the field of molecular biology in the 1980s, and was hailed as another milestone in the development history of molecular biology resources. Once this t...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/686
Inventor 王清水余彦
Owner FUJIAN NORMAL UNIV
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