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Massively parallel sequencing of STRs using a 29‐plex panel reveals stutter sequence characteristics
Author(s) -
Wu Hao,
Ji AiCen,
Liu YiCheng,
Kang KeLai,
Zhang Chi,
Li ZhiWen,
Ji AnQuan,
Ye Jian,
Nie ShengJie,
Wang Le
Publication year - 2020
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.202000093
Subject(s) - massive parallel sequencing , biology , genetics , dna sequencing , microsatellite , amelogenin , locus (genetics) , population , allele , dna , gene , demography , sociology
Massively parallel sequencing of forensic STRs simultaneously provides length‐based genotypes and core repeat sequences as well as flanking sequence variations. Here, we report primer sequences and concentrations of a next‐generation sequencing (NGS)‐based in‐house panel covering 28 autosomal STR loci (CSF1PO, D1GATA113, D1S1627, D1S1656, D1S1677, D2S441, D2S1776, D3S3053, D5S818, D6S474, D6S1017, D6S1043, D8S1179, D9S2157, D10S1435, D11S4463, D13S317, D14S1434, D16S539, D18S51, D18S853, D20S482, D20S1082, D22S1045, FGA, TH01, TPOX, and vWA) and the sex determinant locus Amelogenin. Preliminary evaluation experiments showed that the panel yielded intralocus‐ and interlocus‐balanced sequencing data with a sensitivity as low as 62.5 pg input DNA. A total of 203 individuals from Yunnan Bai population were sequenced with this panel. Comparative forensic genetic analyses showed that sequence‐based matching probability of this 29‐plex panel reached 2.37 × 10 −29 , which was 23 times lower than the length‐based data. Compound stutter sequences of eight STRs were compared with parental alleles. For seven loci, repeat motif insertions or deletions occurred in the longest uninterrupted repeat sequences (LUS). However, LUS and non‐LUS stutters co‐existed in the locus D6S474 with different sequencing depth ratios. These results supplemented our current knowledge of forensic STR stutters, and provided a sound basis for DNA mixture deconvolution.