Primer composition, kit and method for detecting Y-SNP haplogroup based on next-generation sequencing technology and application of primer composition, kit and method
A next-generation sequencing technology, a Y-SNP technology, is applied in biochemical equipment and methods, recombinant DNA technology, and microbial assay/inspection, etc., and can solve problems such as limited technical throughput and inability to detect Y-SNP sites. Achieve the effect of saving time and cost, accurate inference, and detailed division
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Embodiment 1
[0036] This embodiment provides a primer composition and method for high-resolution detection of Y-SNP haplogroups based on next-generation sequencing technology. The steps of specific Y-SNP site screening, primer design and detection method are as follows:
[0037] 1. Y-SNP site screening and primer design;
[0038] According to the proportion of the main haplogroups in the Chinese population, combined with the population of each group, different numbers of downstream branch Y-SNP genetic markers were selected from the Y chromosome phylogenetic tree. In order to ensure the complete structure of the tree, many important node trunk haplogroups are added at the same time. Referring to the 2019 ISOGG website (Version: 15.58) (https: / / isogg.org / tree / HaplogroupTr2019.html), a total of 381 Y-SNP sites for the Chinese population were screened out, forming 359 haplogroups. The detailed information such as the chromosomal position of the selected 381 Y-SNP sites in the human reference g...
Embodiment 2
[0075] This example is the forensic verification of the method for high-resolution detection of Y-SNP haplogroups based on next-generation sequencing technology provided in Example 1.
[0076] In accordance with the requirements of the Scientific Working Group for DNA Analysis Methods (SWGDAM), the sequencing quality, sensitivity, repeatability, accuracy, and applicability of case samples of the method constructed in Example 1 were evaluated and forensic parameters were calculated.
[0077] The average sequencing depth of each Y-SNP site calculated based on 100 samples is as follows figure 2 As shown, the library was constructed smoothly and the sequencing quality was good.
[0078] The results of forensic science verification show that the recommended DNA input amount of this method is at least 1.25ng, but it is still possible to obtain meaningful haplogroup results for micro-sample with extremely small DNA concentration in forensic practice. The sensitivity test results are...
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