Human autosomal STR polymorphic site compound amplification kit and application thereof
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A polymorphic locus, autosomal technology, applied in the field of molecular biology, can solve the problem of less stability than foreign manufacturers, etc.
Active Publication Date: 2018-04-13
AGCU SCIENTECH
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Abstract
Description
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Application Information
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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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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 ...
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 ...
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Abstract
The invention discloses a human autosomal STR polymorphic site compound amplification kit and application thereof. The kit contains specific amplimers for amplifying the following 25 genetic locus: D3S1358, D13S317, D7S820, D16S539, D16S539, D1S1656, PentaE, TPOX, TH01, D2S1338, CSF1PO, Penta D, D19S433, vWA, D21S11, D18S51, D6S1043, Amelogenin, D8S1179, D8S1179, D5S818, D12S391, FGA, D22S1045, Yindel, D2S441 and D10S1248. The kit disclosed by the invention has the advantages that on the premise that the cumulative identification ability of the genetic loci is enhanced, the sensitivity is improved, all the genetic 25 loci can be detected under the condition that the DNA template amount is below 50pg and 29 cycle amplification is carried out, so that a complete genotype can be obtained. According to the human autosomal STR polymorphic site compound amplification kit disclosed by the invention, the quantity of the genetic loci can better satisfy continuous expansion of database construction and detection requirements of the genetic loci are added; the kit is efficient and stable, so that the detection rate of a detection material in a field case is improved and the expansion time isshortened.
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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