STR typing method for loca D8S1179 based on next generation sequencing
A technology of D8S1179-RP and D8S1179, applied in the field of STR typing of locus D8S1179, which can solve the problems of allele loss, limited genetic information, and difficulty in individual identification of trace or complex problems
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Embodiment 1D8
[0031] Embodiment 1D8S1179 Gene Specific Fragment Amplification
[0032] With the consent of the blood donor, normal blood was collected with an anticoagulant tube, and the blood collection tube was immediately placed in an ice box. All blood samples were analyzed by molecular biology method, compared with the human genome sequence reported in NCBI database, using PCR method, according to the primer design principle, using PrimerExpressVersion3 software to design primers in the conserved sequence region of D8S1179 gene:
[0033] D8S1179-FP:5'-TTTGTATTTCATGTGTATCATTCGTATC-3'
[0034] D8S1179-RP:5'ACCTATCCTGTAGATTATTTTCACTGTG-3'
[0035] 1. Use Chelex100 kit to extract DNA from blood of different samples:
[0036] Cut an appropriate amount of blood spots, put them in a centrifuge tube for 0.5 minutes, add appropriate amount of pure water, soak at room temperature, centrifuge at 13,000 rpm for 5 minutes, discard the supernatant, leave about 20 μL of liquid and test material mat...
Embodiment 2
[0063] Example 2 Establishment of a method for analyzing biological information of the D8S1179 gene by next-generation sequencing
[0064] After the library is successfully constructed, take out all the sequencing sequences containing the sample bacode in the original data, first count the length distribution, distinguish the DNA sequence of each sample according to the index sequence in the sample primer, count the number of repeats of the same DNA molecule, and find out the occurrence of SNPs, flanking sequences were retrieved and analyzed. The results shown in Table 2 can appear:
[0065] Table 2
[0066]
[0067] It can be seen that there are 12 and 13 repeats in this sample and in the second repeat, a SNP site with A mutation to G appeared.
[0068] Embodiment 3 Application experiment 1
[0069] For the existing 9 samples of the first-generation typing results, extract DNA, use the primers of the present invention, carry out the PCR method according to the method of...
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