Polymerase chain reaction (PCR) amplification buffer solution for high-GC-content genes, and amplification method and application of PCR amplification buffer solution

Abstract

The invention discloses a polymerase chain reaction (PCR) amplification buffer solution for high-GC-content genes. The PCR amplification buffer solution for the high-GC-content genes is prepared from the following components: 150m mol/L of Tris-HCl, 40 m mol/L of (NH4)2SO4, 2.6 mol/L of betaine monohydrate, 0.02% of emulsifying agent Tween 20 and 20% of PCR reaction enhancer; after the components are collocated, a product is filtered to remove bacteria and then is stored at the temperature of 20 DEG C below zero. The invention also discloses an amplification method and application based on the PCR amplification buffer solution for the high-GC-content genes. The PCR amplification buffer solution uses the (NH4)2SO4 to replace KCL, thus improving the specificity of combination of a primer and a template and being capable of being compatible with a broader spectrum of Mg<2+> range.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a buffer solution for PCR amplification of genes with high GC content and its amplification method and application. Background technique [0002] Polymerase Chain Reaction (PCR) is an in vitro nucleic acid amplification technique developed in the mid-1980s. The PCR process is generally divided into three stages: denaturation, annealing, and extension. First, the DNA is denatured and melted under high temperature conditions. When the temperature drops to about 55°C, the primer binds to the template DNA single strand, and then proceeds under the action of DNA polymerase. DNA synthesis. Compared with the previously commonly used bacteria to amplify target genes, it has the advantages of strong specificity, high sensitivity, simplicity, good repeatability, and easy automation. PCR technology has been developed for nearly 30 years. With the continuous advancement of technolog...

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Problems solved by technology

[0005] In order to solve the technical problem that the above-mentioned existing PCR amplification buffer has little specific effect on the combination of primers and templates, and the chance of amplifying the target band is small, the present invention provides a method that has obvious specific effects on the combination of primers and templates, PCR amplification buffer and amplification method for high GC content gene with high chance of amplifying target band

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