Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot3589

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Generation of Bidirectional Sets of Deletion Mutants by Digestion with BAL 31 Nuclease

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

In this method, the nuclease BAL 31 is used to make uni- or bidirectional deletions in a segment of cloned DNA. BAL 31 is a complex enzyme and tends to digest a population of double-stranded DNA targets in an asynchronous fashion, Deletions created by BAL 31 are therefore far more heterogeneous in size than those created by processive enzymes such as exonuclease III. Warning! BAL 31 is a tricky enzyme. This protocol is not for the technically challenged! Please see the information panel on BAL31 on page 13.68 in the print version of the manual.


MATERIALS

recipe 5x BAL 31 buffer (13-10)

recipe . . . [Full Text of this Article]


METHOD


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