Forensic next-generation sequencing kit based on 74 Y chromosome SNP genetic markers
A technology of Y chromosome and genetic markers, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., to achieve high system resolution, high stability, and accurate typing
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Embodiment 1
[0082] The preparation of embodiment 1 kit of the present invention
[0083] The Y chromosome SNP next-generation sequencing kit for detection may include the following reagents packaged separately:
[0084] a) Multiplex amplification primer mix. From the mixture of amplification primers shown in Table 3, synthesized by Thermo Fisher and mixed into a primer pool.
[0085] b) Compound amplification reaction mixture, partially digested primer sequence reagents. In this example, Thermo Fisher IonAmpliSeq was used TM The PCR reaction mix provided by Library Kit 2.0.
[0086] c) Adapter ligation reagents. In this embodiment, Ion XPress of Thermo Fisher is used TM For BarcodeAdapter, mix adapters and tags 1:1 to form a tag-adapter mixture, dilute 2 times for later use. It should be noted that since the main function of the tag is to distinguish different samples, it is not necessary to use the tag when the number of single sequencing samples is only 1.
[0087] d) Library p...
Embodiment 2
[0092] Example 2 Using the kit of the present invention to detect 100 unrelated Han male individual specimens
[0093] Using the above-mentioned forensic next-generation sequencing kit based on Y chromosome SNP genetic markers, 100 unrelated Han adult male volunteers were tested. The specific detection process is carried out as follows:
[0094] a. DNA was extracted from blood samples of 100 irrelevant Han male individuals by kit or salting-out method, and used as a composite amplification template (the DNA extracted by the salting-out method came from the existing DNA in the laboratory);
[0095] b. Using the DNA template in step a, use the above compound amplification reaction mixture and compound amplification primer mixture to carry out compound amplification of the DNA template and 2800M standard DNA in a single tube to construct each sample library. The system is 20 μL: amplification primer mixture 10 μL, compound amplification reaction mixture 4 μL, template DNA 1 μL, ...
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