Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot2936

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Isolation of DNA Fragments from Polyacrylamide Gels by the Crush and Soak Method

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

The "crush and soak" method, which works best for DNAs <1 kb in size, can be used to recover both single- and double-stranded DNAs from polyacrylamide gels. The yield of eluted DNA varies from <30% to >90% depending on the size of the DNA fragment.


MATERIALS

recipe 6x Gel-loading buffer

recipe Acrylamide gel elution buffer

caution Chloroform

DNA markers generated by restriction digests of known quantities of the DNA sample

DNA sample

Ethanol

Optional, please see Step 5.

caution Phenol:chloroform (1:1, v/v)

recipe caution Sodium acetate (3 M, pH 5.2)

recipe TE (pH 8.0)


METHOD

1. Carry out polyacrylamide gel electrophoresis of the DNA sample and markers as described . . . [Full Text of this Article]


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