Department of Diagnostics and Public Health, Unit of Forensic Medicine - Forensic Genetics Laboratory, University of Verona, Italy.
Med Sci Law. 2021 Jan;61(1_suppl):92-95. doi: 10.1177/0025802420923163.
The introduction of next generation sequencing (NGS; also known as massively parallel sequencing) technology in the field of forensic genetics has been welcomed by the scientific community, above all because it complements the weaknesses of capillary electrophoresis (CE) in the analysis of genetic markers, such as single nucleotide polymorphism (SNP) typing. However, one of the main obstacles to its adoption does not seem to be the cost of the instrumentation, but rather the cost of the NGS library preparation kits. With the aim of reducing the cost of library preparation without compromising the quality of the results, we tried to scale down reaction volumes for the first two polymerase chain reactions in the amplification and enrichment phases of the targeted loci of library preparation using the ForenSeq™ DNA Signature Prep kit. We used 1 µL templated DNA input to a concentration of 1 ng/µL, instead of the 5 µL at 0.2 ng/µL recommended by the manufacturer. Our findings indicate that reduction of the library preparation volume using the ForenSeq™ DNA Signature Prep kit did not interfere with the quality and reproducibility of the DNA profiles obtained and can help lower the overall cost of NGS.
下一代测序（NGS；也称为大规模平行测序）技术在法医遗传学领域的引入受到科学界的欢迎，主要是因为它弥补了毛细管电泳（CE）在分析遗传标记（如单核苷酸多态性（SNP）分型）方面的弱点。然而，采用该技术的主要障碍似乎不是仪器的成本，而是 NGS 文库制备试剂盒的成本。为了在不影响结果质量的情况下降低文库制备成本，我们尝试使用 ForenSeq™ DNA Signature Prep 试剂盒，将文库制备中靶向区域的扩增和富集阶段的前两个聚合酶链反应的反应体积缩小。我们使用浓度为 1ng/µL 的 1µL 模板化 DNA 输入，而不是制造商推荐的 5µL 0.2ng/µL。我们的研究结果表明，使用 ForenSeq™ DNA Signature Prep 试剂盒减少文库制备体积不会干扰获得的 DNA 谱的质量和重现性，并且有助于降低 NGS 的总体成本。
