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用于非Illumina测序平台的降解样本测序文库制备

Sequencing Library Preparation from Degraded Samples for Non-illumina Sequencing Platforms.

作者信息

Martins Renata F, Kampmann Marie-Louise, Förster Daniel W

机构信息

Department of Evolutionary Genetics, Leibniz Institute for Zoo and Wildlife Research (IZW), Berlin, Germany.

Institute for Biochemistry and Biology, University of Potsdam, Potsdam, Germany.

出版信息

Methods Mol Biol. 2019;1963:85-92. doi: 10.1007/978-1-4939-9176-1_10.

Abstract

Efficient methods for building genomic sequencing libraries from degraded DNA have been in place for Illumina sequencing platforms for some years now, but such methods are still lacking for other sequencing platforms. Here, we provide a protocol for building genomic libraries from degraded DNA (archival or ancient sample material) for sequencing on the Ion Torrent™ high-throughput sequencing platforms. In addition to a reduction in time and cost in comparison to commercial kits, this protocol removes purification steps prior to library amplification, an important consideration for work involving historical samples. Libraries prepared using this method are appropriate for either shotgun sequencing or enrichment-based downstream approaches.

摘要

多年来,针对Illumina测序平台,已经有了从降解DNA构建基因组测序文库的有效方法,但其他测序平台仍缺乏此类方法。在此,我们提供了一种从降解DNA(存档或古代样本材料)构建基因组文库的方案,用于在Ion Torrent™高通量测序平台上进行测序。与商业试剂盒相比,该方案不仅减少了时间和成本,还在文库扩增前省去了纯化步骤,这对于涉及历史样本的工作而言是一个重要考量因素。使用此方法制备的文库适用于鸟枪法测序或基于富集的下游方法。

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