An efficient and sensitive method for preparing cDNA libraries from scarce biological samples
Abstract
The preparation and high-throughput sequencing of cDNA libraries from samples of small RNA is a powerful tool to quantify known small RNAs (such as microRNAs) and to discover novel RNA species. Interest in identifying the small RNA repertoire present in tissues and in biofluids has grown substantially with the findings that small RNAs can serve as indicators of biological conditions and disease states. Here we describe a novel and straightforward method to clone cDNA libraries from small quantities of input RNA. This method permits the generation of cDNA libraries from sub-picogram quantities of RNA robustly, efficiently and reproducibly. We demonstrate that the method provides a significant improvement in sensitivity compared to previous cloning methods while maintaining reproducible identification of diverse small RNA species. This method should have widespread applications in a variety of contexts, including biomarker discovery from scarce samples of human tissue or body fluids.Source
Nucleic Acids Res. 2015 Jan 9;43(1):e1. doi: 10.1093/nar/gku637. Epub 2014 Jul 23. Link to article on publisher's siteDOI
10.1093/nar/gku637Permanent Link to this Item
http://hdl.handle.net/20.500.14038/44426PubMed ID
25056322Related Resources
Link to Article in PubMedDistribution License
http://creativecommons.org/licenses/by/4.0/ae974a485f413a2113503eed53cd6c53
10.1093/nar/gku637
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Except where otherwise noted, this item's license is described as http://creativecommons.org/licenses/by/4.0/