Department of Cell Biology and Neuroscience, Institute for Integrative Genome Biology, Center for Disease Vector Research, University of California, Riverside, CA, USA.
Methods Mol Biol. 2021;2170:19-33. doi: 10.1007/978-1-0716-0743-5_2.
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.
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。
