Kit for realizing multiple amplification of 21 short tandem repeats by adopting degenerate primers

Abstract

The invention provides a kit for realizing multiple amplification of 21 short tandem repeats by adopting degenerate primers, and belongs to a multiple amplification system for simultaneously analyzing a plurality of STR gene loci. 21 gene loci are amplified in a multiple amplification system simultaneously, and are divided into the following four groups: the first group: D19S433, TH01, D21S11, D18S51 and Penta E; the second group: D3S1358, D13S317, D7S820, D5S818, CSF1PO and Penta D; the third group: Amelogenin, vWA, D8S1179, TPOX, D6S1043 and D1S1656; and the fourth group: D16S539, D12S391, D2S1338 and FGA. Specific primers are respectively designed on flanks of a gene loci repetitive sequence, so that the balance of the peaks of the fragments inside the gene loci groups achieves 40% or above, the four gene loci groups are subjected to multiple amplification, and the concentration of the primer of each gene locus is regulated, so that the integral balance of the peaks of the gene loci achieves 30% or above. With the kit, a balanced and stable multiple amplification result is easily obtained, so that the kit belongs to a multiple amplification system with low cost and high efficiency.

Description

technical field [0001] The invention relates to the field of biotechnology, and relates to the detection of polymorphic genetic marker genes in human genomes. The present invention relates to the simultaneous amplification of multiple short tandem repeat loci in one amplification system using polymerase chain reaction. Use the method of degenerate primers to improve and increase the amplification efficiency of primer sequences at human single nucleotide polymorphism (SNP) sites, and effectively avoid the phenomenon of locus loss or typing errors caused by SNPs. Background technique [0002] Short tandem repeats (STR for short), also known as microsatellites or simple sequence repeats (SSR), are a type of DNA tandem repeats widely present in eukaryotic genomes, with a core sequence of 2-6 base repeat units. The number of STR loci is large and widely divided, accounting for about 3% of the entire genome (International Human Genome Sequencing Consortium, 2001), and the polymor...

AI Technical Summary

Problems solved by technology

The last base of the primer at the 3' end of the gender recognition site is amplified. Some samples with SNP mutations are prone to amplification failure, resulting in the deletion of this site.

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