Unit

UNIT 4.15 Synthesis and Labeling of RNA In Vitro

  1. Chao Huang1,
  2. Yi-Tao Yu2

Published Online: 1 APR 2013

DOI: 10.1002/0471142727.mb0415s102

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Huang, C. and Yu, Y.-T. 2013. Synthesis and Labeling of RNA In Vitro. Current Protocols in Molecular Biology. 102:4.15.1–4.15.14.

Author Information

  1. 1

    Process Science Downstream, Bristol-Myers Squibb Company, East Syracuse, New York

  2. 2

    Department of Biochemistry and Biophysics, Center for RNA Biology, University of Rochester Medical Center, Rochester, New York

Publication History

  1. Published Online: 1 APR 2013
  2. Published Print: APR 2013

Abstract

This unit discusses several methods for generating large amounts of uniformly labeled, end-labeled, and site-specifically labeled RNAs in vitro. The methods involve a number of experimental procedures, including RNA transcription, 5′ dephosphorylation and rephosphorylation, 3′ terminal nucleotide addition (via ligation), site-specific RNase H cleavage directed by 2′-O-methyl RNA-DNA chimeras, and 2-piece splint ligation. The applications of these RNA radiolabeling approaches are also discussed. Curr. Protoc. Mol. Biol. 102:4.15.1–4.15.14. © 2013 by John Wiley & Sons, Inc.

Keywords:

  • in vitro transcription;
  • radiolabeled RNAs;
  • RNA research