Human autosomal str polymorphic site multiplex amplification kit and its application

A polymorphic locus, autosomal technology, applied in the field of molecular biology, can solve the problem of less stability than foreign manufacturers, and achieve the effect of increasing requirements, improving detection rate, and improving sensitivity

Active Publication Date: 2021-06-29
AGCU SCIENTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, for a long time, domestic manufacturers have not been as stable as foreign manufacturers due to the performance of the kits, such as sensitivity and anti-inhibition ability, and they have only been used in the library building market for a long time, while the actual case inspection materials are all foreign manufacturers’ products. In this area, domestic The application of the kit is blank

Method used

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  • Human autosomal str polymorphic site multiplex amplification kit and its application
  • Human autosomal str polymorphic site multiplex amplification kit and its application
  • Human autosomal str polymorphic site multiplex amplification kit and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] The kit of the present invention consists of the following: primer mixture, 1mL; genomic DNA, 10ng; PCR buffer Super Mix, 2mL; sdH 2 O, 1.95mL; PCR enhancer; Among them, the composition of PCR buffer Super Mix is: Tris-HCl 50mM~125mM with pH8.3~8.5, dNTPs 5.0mM~7.5mM, KCl 50mM~125mM, BSA 2mg / ml~5mg / ml, 5U / μL hot-start Taq enzyme 100μL-300μL; PCR enhancer is trehalose, polyethylene glycol with a concentration lower than 5%, dimethyl sulfoxide, Tween20, NP40, Brij58 or PEG100.

[0043] 1. The operation steps are as follows:

[0044] 1.1 The amplification system is:

[0045] components volume Super Mix 10.0 μL extract sample 6μL The primer of claim 1 5.0 μL wxya 2 o

Make up to 25.0 μL

[0046] 1.2 The amplification procedure is:

[0047]

[0048]

[0049] 1.3 Fluorescence detection of amplified products on a genetic analyzer

[0050] The sample loading mixture is composed of deionized formamide and the molecular we...

Embodiment 2

[0056] 1. Samples such as blood stains, saliva spots, and exfoliated cells in actual cases use the kit of the present invention. Primer Mix, 1mL; Genomic DNA, 10ng; PCR Buffer Super Mix, 2mL; sdH 2 O, 1.95mL; PCR enhancer; Among them, the composition of PCR buffer Super Mix is: Tris-HCl 50mM~125mM with pH8.3~8.5, dNTPs 5.0mM~7.5mM, KCl50mM~125mM, BSA 2mg / ml~5mg / ml, 5U / μL hot-start Taq enzyme 100μL-300μL; PCR enhancer is trehalose, polyethylene glycol with a concentration lower than 5%, dimethyl sulfoxide, Tween20, NP40, Brij58 or PEG100.

[0057] 2. Sample preparation: extract by magnetic bead method, refer to "GA / T 383-2002 Forensic Science DNA Laboratory Inspection Specification" for specific methods.

[0058] 3. The operation steps are as follows:

[0059] 3.1 The amplification system is:

[0060] components volume Super Mix 6.0μL extract sample 1-6μL The primer of claim 1 3.0 μL wxya 2 o

Make up to 15 μL

[0061] 3.2 The...

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Abstract

The invention discloses a human autosomal STR polymorphic site complex amplification kit, the kit includes specific amplification primers for amplifying the following 25 loci; the loci are: D3S1358, D13S317, D7S820, D16S539, D1S1656, PentaE, TPOX, TH01, D2S1338, CSF1PO, Penta D, D19S433, vWA, D21S11, D18S51, D6S1043, Amelogenin, D8S1179, D5S818, D12S391, FGA, D22S1045, Yindel, D22. The kit of the present invention has the following advantages: the kit of the present invention improves the sensitivity under the premise that the identification ability of gene locus accumulation is enhanced, so that 25 loci and 29 loci can be detected under 50 pg of DNA template amount. Cyclic amplification can obtain complete typing; the number of loci in the invention can meet the continuous expansion of database construction and increase the requirements for detection of loci; the kit is efficient and stable, which improves the detection rate of case-site inspection materials and shortens the time of expansion. Add time.

Description

technical field [0001] The invention belongs to the field of molecular biology and relates to STR molecular markers, in particular to a human autosomal STR polymorphic site complex amplification kit covering more loci and higher individual recognition ability and its application. Background technique [0002] Short tandem repeat loci (STR) are a class of DNA sequences formed by tandem repeats of several base pairs as the core unit. The number of core units of tandem repeats is variable, so STR has good genetic polymorphism. Regarding STR, it has been discovered as early as the early 1990s. STR locus fragments are small, can be multiplexed and amplified, have high sensitivity, and can quickly provide accurate and rich information, and have been widely used in the establishment of DNA databases. In 1997, the US FBI selected 13 STR loci: CSF1PO, D16S539, FGA, D18S51, TH01, D5S818, TPOX, VWA, D3S1358, D13S317, D7S820, D8S1179, D21S11; and this year (January 1, 2017 ) The FBI ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888
CPCC12Q1/6888C12Q2600/156C12Q2600/16
Inventor 石美琳姜颖晔范晓芸王伟陈林丽邵泽香郭育林夏子芳郑卫国
Owner AGCU SCIENTECH
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