Retinal Transcriptome Profiling by Directional Next-Generation Sequencing Using 100 ng of Total RNA

  • Matthew J. Brooks
  • Harsha Karur Rajasimha
  • Anand SwaroopEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 884)


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.

Key words

RNA Sequencing Next-generation Sequencing Massively Parallel Sequencing Directional RNA-seq Low input RNA 



The authors are supported by Intramural Research Program of the National Eye Institute, National Institutes of Health, Bethesda, MD, USA.


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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Matthew J. Brooks
    • 1
  • Harsha Karur Rajasimha
    • 1
  • Anand Swaroop
    • 1
    Email author
  1. 1.Neurobiology Neurodegeneration and Repair LaboratoryNational Eye Institute, National Institutes of HealthBethesdaUSA

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