|
|||||
|
|
Massively parallel sequencing of STRs using a 29-plex panel reveals stutter sequence characteristics |
|
| Authors: | Hao Wu Ai-Cen Ji Yi-Cheng Liu Ke-Lai Kang Chi Zhang Zhi-Wen Li An-Quan Ji Jian Ye Sheng-Jie Nie Le Wang |
| Institution: | 1. National Engineering Laboratory for Forensic Science, Key Laboratory 2. of Forensic Genetics of Ministry of Public Security, Institute of Forensic Science, Ministry of Public Security, Beijing, P. R. China
These authors contributed equally to this work.;3. of Forensic Genetics of Ministry of Public Security, Institute of Forensic Science, Ministry of Public Security, Beijing, P. R. China School of Forensic Medicine, Kunming Medical University, Kunming, P. R. China These authors contributed equally to this work.;4. of Forensic Genetics of Ministry of Public Security, Institute of Forensic Science, Ministry of Public Security, Beijing, P. R. China;5. of Forensic Genetics of Ministry of Public Security, Institute of Forensic Science, Ministry of Public Security, Beijing, P. R. China Public Security Bureau of Ximeng County in Pu'er City, Pu'er, P. R. China;6. School of Forensic Medicine, Kunming Medical University, Kunming, P. R. China |
| Abstract: | 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. |
| Keywords: | Forensic genetics Next-generation sequencing STR Stutter sequence Yunnan Bai population |