UNIT 2.10 Hybridization Analysis of DNA Blots
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Molecular Biology
How to Cite
Brown, T. 2001. Hybridization Analysis of DNA Blots. Current Protocols in Molecular Biology. 21:2.10.1–2.10.16.
- Published Online: 1 MAY 2001
- 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 an unsophisticated procedure for hybridization analysis with a radiolabeled DNA probe. The 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.