Method for solving allele amplification imbalance during multiplex PCR through base mismatching

Abstract

The invention discloses a method for solving allele amplification imbalance during multiplex PCR through base mismatching. The method is used for solving the condition that allele amplification imbalance is caused by multiplex PCR. The method comprises the following steps: artificially introducing a mismatching base into a sequence of a primer with relatively high amplification efficiency in original multiplex PCR primers so as to weaken binding power between the primer with relatively high amplification efficiency and a template and then solve the problem of allele amplification imbalance, wherein the original multiplex PCR primers are adopted when the allele amplification imbalance condition occurs during the multiplex PCR amplification of alleles to be assayed. Proven by experiments, the method indeed can be used for effectively solving the allele amplification imbalance caused by the multiplex PCR.

Description

technical field [0001] The invention belongs to the field of gene amplification, and relates to a method for solving the imbalance of allele amplification, in particular to a method for solving the imbalance of allele amplification during multiple PCR by base mismatching. Background technique [0002] PCR technology is based on the polymerase chain reaction, which is a technology that can specifically amplify specific gene fragments, and can increase gene fragments by tens of thousands of times. The core of its principle is that oligonucleotide primers combine and hybridize with a complementary sequence in the single-stranded DNA template to form partial double strands, and DNA synthesis is carried out under the action of DNA polymerase. The combination of oligonucleotide primers and single-stranded DNA templates is based on the principle of base pairing: base pairing follows G (guanine): C (cytosine), A (adenine): T (thymine) / U ( Uracil) based on the Watson-Crick base pa...

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Problems solved by technology

[0005] However, the existing solutions have the following disadvantages: multiplex PCR with universal primers, in the face of highly complex genes, such as HLA genes, because there are a large number of alleles, only conventional multiplex PCR amplification and multiplex PCR combined with universal primers Amplification strategy, not a good solution to the problem of allelic amplification imbalance

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