A method for analyzing the haplotype of PCR products by asynchronous synthesis and sequencing of two nucleotides

Abstract

The invention discloses a method for analyzing PCR product haplotype by synchronous synthesis and sequencing of two nucleotides. Through real-time synthesis and sequencing of two nucleotides, two different nucleotides are added to each sequencing reaction at the same time, and the synthesis and sequencing of different DNA templates in the PCR product are not synchronized, so that the number of different bases on two adjacent SNP sites Information is provided by the different sequencing reactions. Based on the sequencing information at different SNP sites obtained from these asynchronous synthesis sequencing reactions, association analysis was performed to determine the haplotype of the PCR product. The invention broadens the analysis range, is simple to operate, and can reduce the analysis cost; it is suitable for the haplotype analysis of multiple mixed samples, can directly measure the ratio of each haplotype in the mixed sample, and can be used to find, Screening for nucleic acid markers.

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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