Journal
METHODS
Volume 44, Issue 1, Pages 3-12Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymeth.2007.09.009
Keywords
miRNA; small RNA; deep sequencing; small RNA libraries; cDNA libraries
Funding
- Howard Hughes Medical Institute Funding Source: Medline
- NIGMS NIH HHS [P01 GM073047-04, P01 GM073047-019001, P01 GM073047-050001, P01 GM073047-05, P01 GM073047-010001, P01 GM073047-02, P01 GM073047-020001, P01 GM073047-030001, P01 GM073047-03, P01 GM073047, P01 GM073047-01, P01 GM073047-040001] Funding Source: Medline
- NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES [P01GM073047] Funding Source: NIH RePORTER
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Distinct classes of small RNAs, 20-32 nucleotides long, play important regulatory roles for diverse cellular processes. It is therefore important to identify and quantify small RNAs as a function of development, tissue and cell type, in normal and disease states. Here we describe methods to prepare cDNA libraries from pools of small RNAs isolated from organisms, tissues or cells. These methods enable the identification of new members or new classes of small RNAs, and they are also suitable to obtain miRNA expression profiles based on clone count frequencies. This protocol includes the use of new deep sequencing methods (454/Roche and Solexa) to facilitate the characterization of diverse sequence pools of small RNAs. (c) 2007 Elsevier Inc. All rights reserved.
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