Unit

UNIT 6.3 Using DNA Fragments as Probes

  1. William M. Strauss

Published Online: 1 MAY 2001

DOI: 10.1002/0471142727.mb0603s13

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Strauss, W. M. 2001. Using DNA Fragments as Probes. Current Protocols in Molecular Biology. 24:II:6.3:6.3.1–6.3.6.

Author Information

  1. Harvard Medical School, Boston, Massachusetts

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: JAN 1991

Abstract

All hybridization methods depend upon the ability of denatured DNA to reanneal when complementary strands are present in an environment near but below their Tm (melting temperature). A detailed procedure is described in which bacteriophage plaques or bacterial colonies bound to a filter membrane are detected by hybridization with a radioactive probe. Hybridization proceeds on prewet filters placed in a sealable plastic bag. After hybridization the filters are removed from the sealed bag, excess probe is washed off, and the filters are autoradiographed to identify the clones that have hybridized with the probe. An Alternate Protocol differs mainly in that formamide is not used in the hybridization solution.