UNIT 2.10 Hybridization Analysis of DNA Blots

  1. Terry Brown

Published Online: 1 MAY 2001

DOI: 10.1002/0471142727.mb0210s21

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Brown, T. 2001. Hybridization Analysis of DNA Blots. Current Protocols in Molecular Biology. 21:IV:2.10:2.10.1–2.10.16.

Author Information

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

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: JAN 1993


The principle of hybridization analysis is that a single-stranded DNA or RNA molecule of defined sequence (the “probe”) can base-pair to a second DNA or RNA molecule that contains a complementary sequence (the “target”) that has been immobilized on a nitrocellulose and nylon (uncharged and charged) membrane support. The approach taken in this unit is to present as the basic protocol an unsophisticated procedure for hybridization analysis with a radiolabeled DNA probe. The alternate protocol describes a similar method for probing DNA blots with a radiolabeled RNA probe. A support protocol for stripping blots for reprobing is also provided. The commentary describes modifications, including changes to prehybridization, hybridization, and wash solution formulations, and alterations to incubation times and conditions, the latter including a discussion of the wash conditions compatible with different degrees of stringency.