Brooks Matthew J, Rajasimha Harsha Karur, Swaroop Anand
Neurobiology Neurodegeneration and Repair Laboratory, National Eye Institute, National Institutes of Health, Bethesda, MD, USA.
Methods Mol Biol. 2012;884:319-34. doi: 10.1007/978-1-61779-848-1_23.
RNA expression profiles produced by next-generation sequencing (NGS) technology (RNA-seq) allow comprehensive investigation of transcribed sequences within a cell or tissue. RNA-seq is rapidly becoming more cost-effective for transcriptome profiling. However, its usage will expand dramatically if one starts with low amount of RNA and obtains transcript directionality during the analysis. Here, we describe a detailed protocol for the creation of a directional RNA-seq library from 100 ng of starting total RNA.
通过下一代测序（NGS）技术（RNA测序）产生的RNA表达谱，能够对细胞或组织内的转录序列进行全面研究。对于转录组分析而言，RNA测序正迅速变得更具成本效益。然而，如果起始RNA量较低且在分析过程中获得转录本方向性，其应用将大幅扩展。在此，我们描述了一种从100 ng起始总RNA构建定向RNA测序文库的详细方案。
