使用100纳克总RNA通过定向新一代测序进行视网膜转录组分析。

Retinal transcriptome profiling by directional next-generation sequencing using 100 ng of total RNA.

作者信息

Brooks Matthew J, Rajasimha Harsha Karur, Swaroop Anand

机构信息

Neurobiology Neurodegeneration and Repair Laboratory, National Eye Institute, National Institutes of Health, Bethesda, MD, USA.

出版信息

Methods Mol Biol. 2012;884:319-34. doi: 10.1007/978-1-61779-848-1_23.

DOI:10.1007/978-1-61779-848-1_23

PMID:22688717

Abstract

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.

摘要

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。