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.

中文翻译：

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使用29重面板对STR进行大规模平行测序，揭示了口吃序列特征。

法医STR的大规模并行测序可同时提供基于长度的基因型，核心重复序列以及侧翼序列变异。在此，我们报告了基于引物序列和基于下一代测序（NGS）的内部专家组的浓度，涵盖了28个常染色体STR位点（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和vWA）和性别决定因素基因位点。初步评估实验表明，该小组获得了位点内和位点间平衡的测序数据，灵敏度低至62.5 pg输入DNA。用该面板对来自云南白族的203个人进行了测序。⁻²⁹，比基于长度的数据低23倍。将八个STR的复合口吃序列与亲本等位基因进行比较。对于七个基因座，在最长的不间断重复序列（LUS）中发生重复基序插入或缺失。但是，D6S474基因座中的LUS和非LUS口吃并存，且测序深度比不同。这些结果补充了我们目前对法医STR口吃的了解，并为DNA混合物解卷积提供了可靠的基础。