Abstract:
Single-cell sequencing is a fast developing and very promising field; however, it is not commonly used in forensics. The main motivation behind introducing this technology into forensics is to improve mixture deconvolution, especially when a trace consists of the same cell type. Successful studies demonstrate the ability to analyze a mixture by separating single cells and obtaining CE-based STR profiles. This indicates a potential use of the method in other forensic investigations, like forensic DNA phenotyping, in which using mixed traces is not fully recommended. For this study, we collected single-source autopsy blood from which the white cells were first stained and later separated with the DEPArray™ N×T System. Groups of 20, 10, and 5 cells, as well as 20 single cells, were collected and submitted for DNA extraction. Libraries were prepared using the Ion AmpliSeq™ PhenoTrivium Panel, which includes both phenotype (HIrisPlex-S: eye, hair, and skin color) and ancestry-associated SNP-markers. Prior to sequencing, half of the single-cell-based libraries were additionally amplified and purified in order to improve the library concentrations. Ancestry and phenotype analysis resulted in nearly full consensus profiles resulting in correct predictions not only for the cells groups but also for the ten re-amplified single-cell libraries. Our results suggest that sequencing of single cells can be a promising tool used to deconvolute mixed traces submitted for forensic DNA phenotyping.
摘要:
单细胞测序是一个快速发展和非常有前途的领域;然而,它不常用于法医。将这项技术引入取证的主要动机是改进混合反卷积,尤其是当一条轨迹由相同的细胞类型组成时。成功的研究证明了通过分离单细胞和获得基于CE的STR谱来分析混合物的能力。这表明该方法可能用于其他法医调查,比如法医DNA表型鉴定,其中不完全建议使用混合痕迹。对于这项研究,我们收集了单一来源的尸检血液,首先对白细胞进行了染色,然后用DEPArray™N×T系统进行了分离.20、10和5个细胞的组,以及20个单细胞,收集并提交DNA提取。使用IonAmpliSeq™PhenoTrivium面板制备文库,其中包括两种表型(HIrisPlex-S:eye,头发,和肤色)和祖先相关的SNP标记。在测序之前,另外扩增和纯化了一半的基于单细胞的文库,以提高文库浓度.祖先和表型分析导致几乎完全的共识谱,从而不仅对细胞组而且对十个重新扩增的单细胞文库都产生了正确的预测。我们的结果表明,单细胞测序可能是一种有前途的工具,用于对提交给法医DNA表型的混合痕迹进行去卷积。