A method for primer selection, sequencing by synthesis, and analysis of haplotypes of PCR products

Abstract

The invention provides a method for analyzing haplotype of PCR products by primer selection sequencing-by-synthesis. The method is adopted to perform haplotype analysis on two or more PCR products (multiple DNA templates) of site variation (such as SNP and mutation), wherein the PCR products are hybridized by a specific sequencing primer and then are divided into two parts, and a 3' terminus-closed monomer reagent is added to each part; the sequencing primer extends one base with the varied site under the guidance of a DNA template, the 3' terminus of the sequencing primer is closed and cannot extend continuously, and then the sequencing-by-synthesis is performed on the unclosed primers to obtain the sequence information of a specific DNA template, thereby determining the haplotype of PCR products.

Description

technical field

[0001] The invention relates to the field of biotechnology, in particular to a method for analyzing PCR product haplotypes by using primers to select, synthesize and sequence. Background technique

[0002] In molecular biology research, DNA sequence analysis is the basis for further research and modification of target genes, and is crucial to life science. With the development and completion of the Human Genome Project and various model organism genome projects, human beings have entered the post-gene era, and the related disciplines of molecular biology have developed rapidly. It will become possible to understand the differences of life, the law of disease occurrence and development, and the interaction between drugs and living organisms at the genetic level. In terms of basic research, research on the inheritance rules of disease genes and clone disease-causing genes; in terms of application, directly find the mutation sites of susceptibility genes of dis...

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