Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot3938

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Lysing Colonies and Binding of DNA to Filters

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 15% of the full text of this article appears below.


INTRODUCTION

In this protocol, based on the procedure of Grunstein and Hogness (1975), alkali is used to liberate DNA from bacterial colonies on nitrocellulose or nylon filters. The DNA is then fixed to the filter by UV-cross-linking or baking under vacuum.


MATERIALS

recipe 2X SSPE

E. coli transformants immoblized on filters

Use transformants prepared by one of the methods described in Screening Bacterial Colonies by Hybridization: Small Numbers, Screening Bacterial Colonies by Hybridization: Intermediate Numbersand Screening . . . [Full Text of this Article]


METHOD


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