Y-SNP heredity labelled forensic medicine composite detection kit based on 20 haplogroups D
A Y-SNP, composite detection technology, applied in the field of forensic composite detection kits, can solve the problem of inability to be assigned to haplogroup D twigs, etc., achieve good group specificity and geographical specificity, wide application and promotion value, Accurate and reliable typing results
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Embodiment 1
[0082] The preparation of embodiment 1 kit of the present invention
[0083] The SNP genetic marker composite detection kit for 20 Y chromosome haplogroup D for detection may include the following reagents packaged separately:
[0084] a) Multiplex amplification primer mix. It was obtained by mixing the amplification primers shown in Table 2. The amplification primers were synthesized by Invitrogen Company. The synthetic amplification primers were prepared with ultrapure water to 100pM / μL, and then mixed according to the ratio in Table 6 to make a composite amplification primer. Primer mix.
[0085] b) Compound amplification reaction mixture. The PCR reaction mixture MultiplexPCRMix of Qiagen Company was used in this embodiment.
[0086] c) Multiple single base extension reaction primer mix. It was obtained by mixing the single-base extension reaction primers shown in Table 3, and the single-base extension reaction primers were all synthesized by Invitrogen Company. The s...
Embodiment 2
[0095] Example 2 Using the kit of the present invention to detect 200 unrelated individual samples roughly classified into haplogroup D
[0096] Using the above-mentioned forensic science composite detection kit based on 20 Y-SNP genetic markers of haplogroup D, 200 samples of unrelated individuals roughly classified as haplogroup D were tested. The specific detection process is as follows:
[0097] a. The DNA of 200 samples was extracted by the Chelex-100 method as a composite amplification template;
[0098] b. Using the DNA in step a as a template, use the composite amplification primer mixture and the composite amplification reaction mixture to perform composite PCR amplification on the sample in the following amplification system; 0.3 μL of the composite amplification primer mixture, the composite amplification reaction Mixture 2.5 μL, template DNA 0.8 μL, add ddH2O to 5 μL; PCR amplification conditions: 95°C, 15 minutes; 94°C, 30 seconds; 66°C-60°C, 90 seconds; 72°C, 30...
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