Appendix

APPENDIX 3K Analysis of RNA by Northern Blot Hybridization

  1. Terry Brown1,
  2. Karol Mackey2

Published Online: 1 NOV 2001

DOI: 10.1002/0471142905.hga03ks30

Current Protocols in Human Genetics

Current Protocols in Human Genetics

How to Cite

Brown, T. and Mackey, K. 2001. Analysis of RNA by Northern Blot Hybridization. Current Protocols in Human Genetics. 30:3K:A.3K.1–A.3K.12.

Author Information

  1. 1

    University of Manchester Institute of Science and Technology, Manchester, United Kingdom

  2. 2

    Molecular Research, Cincinnati, Ohio

Publication History

  1. Published Online: 1 NOV 2001
  2. Published Print: AUG 2001

Abstract

Specific sequences in RNA preparations can be detected by blotting and hybridization analysis using techniques very similar to those originally developed for DNA. Fractionated RNA is transferred from an agarose gel to a membrane support (northern blotting); unfractionated RNA is immobilized by slot or dot blotting. The resulting blots are studied by hybridization analysis with labeled DNA or RNA probes. Because they are single-stranded, most RNAs are able to form secondary structures by intramolecular base pairing and must therefore be electrophoresed under denaturing conditions if good separations are to be obtained. The Basic Protocol describes blotting and hybridization of RNA fractionated in an agarose-formaldehyde gel. Alternate protocols describe the glyoxal/DMSO method for denaturing gel electrophoresis and slot-blot hybridization of RNA samples. Stripping hybridization probes from blots can be done under three different sets of conditions; these methods are outlined in a Support Protocol.