Method for solving allele amplification unbalance by PCR additive

An allele amplification and allele technology, which is applied in the field of unbalanced allelic amplification and can solve problems such as unbalanced allelic amplification

Inactive Publication Date: 2017-07-25
BOAO BIOLOGICAL CO LTD +1
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  • Description
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Problems solved by technology

[0006] In order to effectively solve the problem of unbalanced allelic amplification,

Method used

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  • Method for solving allele amplification unbalance by PCR additive
  • Method for solving allele amplification unbalance by PCR additive
  • Method for solving allele amplification unbalance by PCR additive

Examples

Experimental program
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Embodiment

[0026] Example, Resolving Allelic Amplification Imbalances by PCR Additives

[0027] PCR additive is a kind of synergist in PCR reaction, common types include formamide, betaine, DMSO, etc. The present invention finds that the PCR additive can reduce the stability of the double-helix DNA, make the secondary structure easy to open, and help the DNA polymerase to extend through the secondary structure region. During the PCR process, the single strand of one allele of the two alleles forms a complex secondary structure or the DNA double strands melt insufficiently, resulting in inconsistent amplification efficiencies of the two alleles, that is, the problem of unbalanced amplification , You can add a certain concentration of betaine-like PCR additives to solve such problems.

[0028] The method for solving the problem of unbalanced allelic amplification by using PCR additives will be described below with specific examples.

[0029] Alleles: HLA-DQB1 genes whose genotypes are DQ...

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Abstract

The invention discloses a method for solving the imbalance of allele amplification through PCR additives. This method is aimed at the situation of unbalanced allelic amplification caused by "some alleles to be tested melt to form complex secondary structures during PCR amplification or/and DNA double-strand melts insufficiently". The method comprises the steps of: reducing the stability of double-helix DNA by adding PCR additives to the original PCR reaction system, thereby solving the problem of unbalanced allelic amplification; the original PCR reaction system is the allele to be tested The PCR reaction system used when the amplification of alleles is unbalanced in the PCR amplification of genes. Experiments have proved that this method can effectively solve the allele amplification caused by "part of the alleles to be tested melted during the PCR amplification process to form a complex secondary structure or/and DNA double-strand melts insufficiently" unbalanced.

Description

technical field [0001] The invention belongs to the field of gene amplification, and relates to a method for solving the unbalanced allelic amplification, in particular to a method for solving the unbalanced allelic amplification by using PCR additives. 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 pairing principle. ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6858
Inventor 刘湘兰更欣赵晓丹王雷陈璐秦雪菲李锦绣
Owner BOAO BIOLOGICAL CO LTD
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