Composite amplification system based on Y-STR locus and specific primer combination thereof
A compound amplification system and primer combination technology, applied in the field of forensic science, can solve the problems that different individuals cannot be distinguished well, the mutation rate of Y-STR loci is not high, and the application of Y-STR loci is limited.
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Embodiment 1
[0122] Embodiment 1, the preparation of the compound amplification system based on Y-STR locus
[0123] 1. Screening of Y-STR loci
[0124] The inventor of the present invention obtains a large amount of candidate multi-copy Y-STR loci by consulting the literature; Then, using the research results of the Human Genome Project, using the reported The primer information of each locus was retrieved to obtain the corresponding sequence; the sequence was retrieved using BLAST (URL: https: / / blast.ncbi.nlm.nih.gov / Blast.cgi), and the existing multi-copy Y - Sequence information of the core and flanking regions of the STR locus, allele length and distribution, etc. were sorted out and systematically studied, focusing on multi-copy Y-STR loci with a core sequence of 4-5 base repeats. Based on technical indicators such as copy number, mutation rate, repeat unit characteristics, and sequence structure complexity of each locus, 35 candidate multi-copy Y-STR loci were initially screened ou...
Embodiment 2
[0135] Embodiment 2, the application of the compound amplification system based on Y-STR locus
[0136] The sample to be tested is sample 1 or sample 2:
[0137] Sample 1, 1ng of human (known as male) genomic DNA;
[0138] Sample 2: Take a blood collection card from a person (known to be a male), and use a 0.5mm hand-held punch to punch out a disc.
[0139] The blood collection card of the person (known as a male) was provided by the Daily DNA Database Construction Institute of the Physical Evidence Identification Center of the Ministry of Public Security.
[0140] 1. Take 10 μL of the compound amplification system based on the Y-STR locus prepared in Step 3 of Example 1, add the sample to be tested for PCR amplification, and obtain the PCR amplification product.
[0141] Reaction program: pre-denaturation at 95°C for 11 min; denaturation at 94°C for 30 s, annealing at 59°C for 2 min, extension at 72°C for 1 min, 28 cycles; extension at 60°C for 60 min; storage at 4°C.
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