Method for analyzing haplotype of PCR product employing non-synchronous synthesis sequencing of two nucleotides

Abstract

The invention discloses a method for analyzing haplotype of a PCR product employing non-synchronous synthesis sequencing of two nucleotides. Through real-time synthesis sequencing of two nucleotides, two different nucleotides are simultaneously added for each sequencing reaction; the synthesis sequencing of different DNA templates in the PCR product is not synchronous, so that information of different base groups on two adjacent SNP sites is provided by different sequencing reactions; and association analysis is carried out according to sequencing information on different SNP sites obtained by these non-synchronous synthesis sequencing reactions, so as to determine the haplotype of the PCR product. The analysis range is expanded; the operation is simple; the analysis cost can be reduced; the method is suitable for haplotype analysis of a plurality of mixing samples; the ratio of individual haplotypes in the mixing samples can be directly determined; and ucleic acid markers can be searched and screened from large samples.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a PCR product haplotype analysis method, in particular to a method for asynchronous synthesis and sequencing of two nucleotides to analyze the PCR product haplotype. Background technique [0002] The Human Genome Haplotype Map Project itself illustrates the importance of haplotype research and also provides an important database for haplotype analysis of individual samples. The combination of a group of SNPs located in a specific region of a chromosome that are related and tend to be inherited as a whole is called a haplotype. Early research work has shown that haplotype analysis can greatly reduce the workload of disease association studies, and provide a larger amount of information than SNP genotype analysis, which is conducive to discovering more information. Theoretically, there may be 2n haplotypes in a SNP region containing N independent biallelic genes, but in practice, due to...

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Problems solved by technology

However, the invention patent requires two sequencing of a DNA template sample, which not only increases the complexity of the operation, but also increases the cost of the analysis, and the information of the simultaneous sequencing reaction cannot meet the analysis of the haplotype

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