Unit

UNIT 2.9A Southern Blotting

  1. Terry Brown

Published Online: 1 MAY 2001

DOI: 10.1002/0471142727.mb0209as21

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Brown, T. 2001. Southern Blotting. Current Protocols in Molecular Biology. 00:IV:2.9A:2.9.1–2.9.20.

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

Abstract

Southern blotting is the transfer of DNA fragments from an electrophoresis gel to a membrane support (the properties and advantages of the different types of membrane, transfer buffer, and transfer method are discussed in detail), resulting in immobilization of the DNA fragments, so the membrane carries a semipermanent reproduction of the banding pattern of the gel. After immobilization, the DNA can be subjected to hybridization analysis, enabling bands with sequence similarity to a labeled probe to be identified. This unit presents several protocols for DNA transfer to nylon and nitrocellulose membranes, and for calibrating a UV transilluminator to cross-link the transferred DNA to the membrane.