Ponts Nadia, Chung Duk-Won D, Le Roch Karine G
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. 2012;883:59-73. doi: 10.1007/978-1-61779-839-9_4.
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 level-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.
仅在过去几年中，新一代测序技术才得以应用，并且迅速衍生出了许多应用，例如RNA测序（RNA-seq）技术的发展，该技术使用深度测序来分析整个转录组。RNA测序有能力发现新的转录本和剪接变体、单核苷酸变异、融合基因以及基于mRNA水平的表达谱。制备RNA测序文库可能很精细，通常需要购买昂贵的试剂盒，而这些试剂盒需要大量起始材料。这里介绍的方法灵活且具有成本效益。使用这种方法，我们从人类疟原虫恶性疟原虫提取的RNA中制备了高质量的链特异性RNA测序文库。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用于其他平台。
