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链特异性 RNA-Seq 在疟疾样本中的应用。

Strand-Specific RNA-Seq Applied to Malaria Samples.

机构信息

Department of Cell Biology and Neuroscience, Institute for Integrative Genome Biology, Center for Disease Vector Research, University of California, Riverside, CA, USA.

出版信息

Methods Mol Biol. 2021;2170:19-33. doi: 10.1007/978-1-0716-0743-5_2.

Abstract

Over the past few years only, next-generation sequencing technologies became accessible and many applications were rapidly derived, such as the development of RNA-seq, a technique that uses deep sequencing to profile whole transcriptomes. RNA-seq has the power to discover new transcripts and splicing variants, single nucleotide variations, fusion genes, and mRNA levels-based expression profiles. Preparing RNA-seq libraries can be delicate and usually obligates buying expensive kits that require large amounts of stating materials. The method presented here is flexible and cost-effective. Using this method, we prepared high-quality strand-specific RNA-seq libraries from RNA extracted from the human malaria parasite Plasmodium falciparum. The libraries are compatible with Illumina's sequencers Genome Analyzer and Hi-Seq. The method can however be easily adapted to other platforms.

摘要

在过去的几年中,下一代测序技术变得可及,并且迅速衍生出许多应用,例如 RNA-seq 的发展,这是一种使用深度测序来描绘整个转录组的技术。RNA-seq 具有发现新转录本和剪接变体、单核苷酸变异、融合基因以及基于 mRNA 水平的表达谱的能力。准备 RNA-seq 文库可能很精细,通常需要购买昂贵的试剂盒,这些试剂盒需要大量的起始材料。这里介绍的方法灵活且具有成本效益。使用这种方法,我们从人类疟疾寄生虫疟原虫提取的 RNA 中制备了高质量的链特异性 RNA-seq 文库。这些文库与 Illumina 的测序仪 Genome Analyzer 和 Hi-Seq 兼容。然而,该方法可以很容易地适应其他平台。

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