Small RNA Library Construction for High-Throughput Sequencing

  • Jon McGinn
  • Benjamin Czech
Part of the Methods in Molecular Biology book series (MIMB, volume 1093)


Since their discovery about 20 years ago, small RNAs have been shown to play a critical role in a myriad of biological processes. The greater availability of high-throughput sequencing has been invaluable to furthering our understanding of small RNAs as regulatory molecules. In particular, these sequencing technologies have been crucial in understanding the role of small RNAs in reproductive tissues, where millions of individual sequences are generated. In this context, high-throughput sequencing provides the requisite level of resolution that other procedures, like northern blotting, would not be able to achieve. Here, we describe a protocol for the preparation of small RNA libraries for sequencing using the Solexa/Illumina technology.

Key words

Endo-siRNA miRNAs piRNAs Small RNA libraries Cloning High-throughput sequencing 



We thank past and current members of the Hannon laboratory that helped improve this protocol, especially Alexei Aravin, Julius Brennecke, and Colin Malone.


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Copyright information

© Springer Science+Business Media, LLC 2014

Authors and Affiliations

  • Jon McGinn
    • 1
  • Benjamin Czech
    • 1
  1. 1.Cold Spring Harbor LaboratoryCold Spring HarborUSA

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