Section of Forensic Genetics, Department of Forensic Medicine, Faculty of Health & Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
Biotechniques. 2020 Jun;68(6):342-344. doi: 10.2144/btn-2019-0156. Epub 2020 Mar 6.
Here, we present the development of an automated AmpliSeq™ (ThermoFischer, MA, USA) workflow for library building using the Biomek 3000 Laboratory Automation Workstation (Beckman Coulter Inc., CA, USA), in which the total volume of PCR reagents and reagents for library preparation are reduced by one-half. The automated AmpliSeq workflow was tested using 43 stain samples (blood, bone, muscle tissue, semen, swab, nail scrape and cigarette butts) collected from crime scenes. The sequencing data were evaluated for locus balance, heterozygous allele balance and noise. The performance of libraries built with the automated AmpliSeq workflow using one-half of the recommended reagent volumes were similar to the performance of libraries built with the recommended (full) volumes of the reagents.
在这里，我们展示了一种自动化的 AmpliSeq（ThermoFischer，MA，USA）文库构建工作流程的开发，该流程使用 Biomek 3000 实验室自动化工作站（Beckman Coulter Inc.，CA，USA），其中 PCR 试剂和文库制备试剂的总体积减少了一半。该自动化 AmpliSeq 工作流程使用从犯罪现场收集的 43 个染色样本（血液、骨骼、肌肉组织、精液、拭子、指甲刮屑和烟蒂）进行了测试。对测序数据进行了定位平衡、杂合等位基因平衡和噪声评估。使用推荐试剂体积的一半构建的自动化 AmpliSeq 文库的性能与使用推荐（全）试剂体积构建的文库的性能相似。
