Max Planck Institute for Molecular Genetics, Ihnestr. 73, D-14195 Berlin, Germany.
Biochem Biophys Res Commun. 2012 Jun 15;422(4):643-6. doi: 10.1016/j.bbrc.2012.05.043. Epub 2012 May 15.
Preserving the original RNA orientation information in RNA-Sequencing (RNA-Seq) experiment is essential to the analysis and understanding of the complexity of mammalian transcriptomes. We describe herein a simple, robust, and time-effective protocol for generating strand-specific RNA-seq libraries suited for the Illumina sequencing platform. We modified the Illumina TruSeq RNA sample preparation by implementing the strand specificity feature using the dUTP method. This protocol uses low amounts of starting material and allows a fast processing within two days. It can be easily implemented and requires only few additional reagents to the original Illumina kit.
在 RNA 测序 (RNA-Seq) 实验中保留原始 RNA 方向信息对于分析和理解哺乳动物转录组的复杂性至关重要。本文描述了一种简单、稳健且高效的方法，用于生成适用于 Illumina 测序平台的链特异性 RNA-seq 文库。我们通过使用 dUTP 方法实现 Illumina TruSeq RNA 样品制备中的链特异性特征来修改该方法。该方案使用少量起始材料，并允许在两天内快速处理。它可以很容易地实现，并且只需要向原始 Illumina 试剂盒添加少量额外的试剂。
