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Rice Protocols pp 109-118 | Cite as

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

  • Guru Jagadeeswaran
  • Ramanjulu SunkarEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 956)

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.

Key words

MciroRNAs Small RNAs Posttranscriptional gene regulation Rice 

Notes

Acknowledgments

This research was supported by the Oklahoma Agricultural Experiment Station and the USDA/NRI grant (2007-02019).

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Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  1. 1.Department of Biochemistry and Molecular BiologyOklahoma State UniversityStillwaterUSA

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