Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4043

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Southern Blotting: Simultaneous Transfer of DNA from a Single Agarose Gel to Two Membranes

Joseph Sambrook and David W. Russell

This protocol was adapted from Molecular Cloning, 3rd edition, by Joseph Sambrook and David W. Russell. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2001

The first 100 words of the full text of this article appear below.


INTRODUCTION

DNA can be simultaneously transferred from opposite sides of a single agarose gel to two membranes. Bidirectional transfer occurs rapidly at first, but soon slows down as the gel becomes dehydrated. Because the efficiency of transfer is low, the method works best when the target sequences are present in high concentration, for example, when analyzing restriction digests of cloned DNAs or less complex genomes.


MATERIALS

recipe 6X SSC

6x SSC should be warmed to various temperatures. Please see Steps 11, 12, and 13.

recipe 6x Gel-loading buffer IV

recipe DNA-staining solution

recipe Denaturation solution

recipe caution HCl (0.2 N), for depurination of DNA

Optional, please see note . . . [Full Text of this Article]


METHOD


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