Laboratory of Pharmaceutical Biotechnology, Ghent University, 9000 Gent, Belgium; Department of Life Sciences and Imaging, imec, 3001 Leuven, Belgium.
Laboratory of Pharmaceutical Biotechnology, Ghent University, 9000 Gent, Belgium.
Forensic Sci Int Genet. 2019 Jan;38:204-210. doi: 10.1016/j.fsigen.2018.11.012. Epub 2018 Nov 12.
The potential and current state-of-the-art of forensic SNP genotyping using nanopore sequencing was investigated with a panel of 16 tri-allelic single nucleotide polymorphisms (SNPs), multiplexing five samples per sequencing run. The sample set consisted of three single-source human genomic reference control DNA samples and two GEDNAP samples, simulating casework samples. The primers for the multiplex SNP-loci PCR were taken from a study which researched their value in a forensic setting using conventional single-base extension technology. Workflows for multiplexed Oxford Nanopore Technologies' 1D and 1D sequencing were developed that provide correct genotyping of most SNP loci. Loci that are problematic for nanopore sequencing were characterized. When such loci are avoided, nanopore sequencing of forensic tri-allelic SNPs is technically feasible.
使用纳米孔测序对法医 SNP 基因分型的潜力和现状进行了研究，使用了一个包含 16 个三等位单核苷酸多态性 (SNP) 的面板，每个测序运行中多重化五个样本。样本集包括三个单源人类基因组参考对照 DNA 样本和两个 GEDNAP 样本，模拟案例样本。用于多重 SNP 基因座 PCR 的引物来自一项研究，该研究使用传统的单碱基延伸技术研究了它们在法医环境中的价值。开发了用于多重 Oxford Nanopore Technologies' 1D 和 1D 测序的工作流程，这些工作流程可提供大多数 SNP 基因座的正确基因分型。对纳米孔测序有问题的基因座进行了特征描述。当避免这些基因座时，法医三等位 SNP 的纳米孔测序在技术上是可行的。
