Retinal transcriptome profiling by directional next-generation sequencing using 100 ng of total RNA.
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.
Neurobiology Neurodegeneration and Repair Laboratory, National Eye Institute, National Institutes of Health, Bethesda, MD, USA.
Matthew J
MJ
Brooks
Harsha Karur
HK
Rajasimha
Anand
A
Swaroop
D000818:Animals; D020869:Gene Expression Profiling / Q000379:methods*; D015723:Gene Library; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D051379:Mice; D012313:RNA / Q000737:chemistry / Q000302:isolation & purification; D012160:Retina / Q000378:metabolism*; D017423:Sequence Analysis, RNA / Q000379:methods*; D059467:Transcriptome
D012313:RNA