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

A technology of compound amplification and compound amplification system, which is applied in the direction of microbial measurement/inspection, biochemical equipment and methods, etc., can solve the problems of easy amplification failure and missing, and achieve good balance, high specificity and sensitivity high effect

Inactive Publication Date: 2016-03-02
BEIJING PEOPLESPOT TECH
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Problems solved by technology

The last base of the primer at the 3' end of the gender recognition site is amplified. Some sampl

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

Examples

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Embodiment 1

[0066] Merger Primer 21 Gene Loci Multiplex Amplification Kit Detects Genotyping of Sample Loci

[0067]Blood is donated by volunteers. Template DNA was extracted from blood by chelex-100 method. The amplification reaction was carried out on the ABI9700 thermal cycler, the electrophoresis and detection were carried out on the ABI3100 genetic analyzer, and the data analysis was performed using GeneMapperIDv3.2 software. The samples were detected by the PP21 kit of Promega Company and the Goldeneye20A kit of Genepoint Cognitive Co., Ltd., and the operation method was carried out according to the instructions of the kits, and the results were used as a control. The reagents and materials used in the present invention, such as allelic ladder (ladder), are conventional materials commonly used by those skilled in the art.

[0068] 1.1. chelex-100 method to extract DNA (specific method refer to "ForensicDNA Protocol", HumanaPress, 1998)

[0069] 1) Take 3 μl of blood with anticoag...

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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...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6858C12Q2525/151C12Q2531/113C12Q2537/143C12Q2563/107
Inventor 张威张琰
Owner BEIJING PEOPLESPOT TECH
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