Forensic medicine compound detection kit based on Y chromosome SNP (single nucleotide polymorphism) genetic marker
A genetic marker, Y chromosome technology, applied in the field of forensic genetics, can solve the problems of difficult selection of Y chromosome SNP genetic markers, limited number of Y-SNPs, and no Y-SNP composite detection system, and achieves widespread promotion and application value. , accurate typing, high-efficiency effect
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Embodiment 1
[0067] Embodiment 1 Preparation of kit of the present invention
[0068] The Y chromosome SNP composite detection kit for detection may include the following reagents packaged separately:
[0069] a) Multiplex amplification primer mix. It was obtained by mixing the amplification primers shown in Table 1, and was synthesized by TaKaRa Biotechnology Company. The synthesized 20 pairs of amplification primers were prepared with ultrapure water to 50pM / μL, and then mixed according to the ratio in Table 6 to make a composite amplification primer. Primer mix.
[0070] b) Compound amplification reaction mixture. The PCR reaction mixture One shot La PCR of TaKaRa Biotechnology company is used in this embodiment TM Mix.
[0071] c) Primer mix for multiplex single base extension reactions. It was obtained by mixing the primers for the single-base extension reaction shown in Table 2, and was synthesized by TaKaRa Biotechnology Company. The synthesized 20 single-base extension react...
Embodiment 2
[0081] Example 2 Using the method of the present invention to detect unrelated Han nationality individual specimens
[0082] Use the kit in Example 1 to detect unrelated Han individuals.
[0083] a. Genomic DNA was extracted from blood samples of 220 unrelated Han individuals by the Chelex-100 method, and used as a template for multiplex amplification.
[0084] b. Using the DNA template in step a, use the composite amplification primer mixture and the composite amplification reaction mixture to perform composite PCR amplification on the sample DNA in the following amplification system.
[0085]
[0086]
[0087] Amplified Thermal Cycling Parameters
[0088]
[0089] Cycle from step 2 to step 4 34 times
[0090] 5. 72°C for 10 minutes
[0091] c. Purification of multiple PCR products, the following is the purification system of each sample amplification product
[0092]
[0093] Amplified product purification reaction conditions:
[0094] 37°C 60 minutes
[00...
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