Forensic medicine composite detection reagent kit based on 20 triallelic SNP genetic markers and detection method
A tri-allelic and genetic marker technology is applied in the field of forensic compound detection kits for allele SNP genetic markers. It can solve the problems of large gaps in genetic markers, limited number of tri-allelic SNPs, and inability to analyze mixed samples. The effect of widespread promotion and application of value
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Embodiment 1
[0059] Embodiment one: the preparation of kit of the present invention
[0060] The tri-allelic SNP composite detection kit for detection may include the following reagents packaged separately:
[0061] A. Multiplex amplification primer mix. It is obtained by mixing the amplification primers shown in Table 1. The synthesized 20 pairs of amplification primers are dissolved in water and mixed according to the concentrations in Table 6.
[0062] B. Compound amplification reaction mixture. The PCR reaction mixture OneshotLaPCR of TaKaRaBiotechnology company is used in this embodiment TM Mix.
[0063] C. Multiple single base extension reaction primer mixture. It is obtained by mixing the single-base extension reaction primers shown in Table 2. The synthesized 20 single-base extension reaction primers are dissolved in water and mixed according to the concentrations in Table 7.
[0064] D. Single base extension reaction mixture. Use ABI company's SNaPshotreadyreactionmix single...
Embodiment 2
[0074] Embodiment two: using the method of the present invention to identify 100 irrelevant Han individual test materials
[0075] Using the above-mentioned forensic medicine review test mixture based on the three-allelic SNP genetic marker, we conducted the test of 100 unrelated individual samples, and the specific identification process was as follows:
[0076] A. Genomic DNA was extracted from blood samples of 100 unrelated Han individuals, and the genomic DNA was obtained as a template.
[0077] B. Prepare the amplification primer pools in each group of single-tube multiplex amplification systems respectively, dilute 20 pairs of amplification primers to 50pM / μL with ultrapure water, mix them according to the ratio in Table 6, and use them as primer pools, where each The final concentrations of the primers are shown in Table 6.
[0078] Using the DNA template in step A, use the primer pool and the multiplex amplification reaction mixture in step B to perform multiplex PCR ...
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