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Cloning of Small RNAs for the Discovery of Novel MicroRNAs in Plants

Authors :
Ramanjulu Sunkar
Guru Jagadeeswaran
Source :
Methods in Molecular Biology ISBN: 9781627031936
Publication Year :
2012
Publisher :
Humana Press, 2012.

Abstract

Endogenous small RNAs can be grouped into several distinct classes of 21-nt-long microRNAs (miRNAs), short interfering RNAs (siRNAs), trans-acting siRNAs (tasiRNAs), and 24-nt long heterochromatic siRNAs. miRNAs are increasingly being recognized as significant effectors of gene regulation in a wide range of organisms. These molecules are typically ∼21-nt long and are amenable for cloning by streamlined protocols. Here we detail the methodology for cloning small RNAs in rice to identify novel miRNAs and other important small RNAs. Briefly, small RNA molecules are size fractionated, attached to adaptors, and subsequently converted into cDNA and PCR amplified. Current high-throughput sequencing technologies allow sequencing of the PCR products directly.

Details

ISBN :
978-1-62703-193-6
ISBNs :
9781627031936
Database :
OpenAIRE
Journal :
Methods in Molecular Biology ISBN: 9781627031936
Accession number :
edsair.doi...........ebdf36cd5da320733bccfc28f3a15d82
Full Text :
https://doi.org/10.1007/978-1-62703-194-3_9