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Methods. 2012 Oct;58(2):164-70. doi: 10.1016/j.ymeth.2012.07.030. Epub 2012 Aug 7.

Barcoded cDNA library preparation for small RNA profiling by next-generation sequencing.

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  • 1Laboratory of RNA Molecular Biology, Howard Hughes Medical Institute, The Rockefeller University, 1230 York Avenue, Box 186, NY 10065, USA.

Abstract

The characterization of post-transcriptional gene regulation by small regulatory (20-30 nt) RNAs, particularly miRNAs and piRNAs, has become a major focus of research in recent years. A prerequisite for characterizing small RNAs is their identification and quantification across different developmental stages, and in normal and disease tissues, as well as model cell lines. Here we present a step-by-step protocol for generating barcoded small RNA cDNA libraries compatible with Illumina HiSeq sequencing, thereby facilitating miRNA and other small RNA profiling of large sample collections.

Copyright © 2012 Elsevier Inc. All rights reserved.

PMID:
22885844
[PubMed - indexed for MEDLINE]
PMCID:
PMC3508525
Free PMC Article
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