Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot3919

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protocolProtocol

Directional Cloning into Plasmid Vectors

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

Directional cloning requires that the plasmid vector be cleaved with two restriction enzymes that generate incompatible termini and that the fragment of DNA to be cloned carries termini that are compatible with those of the doubly cleaved vector.


MATERIALS

recipe ATP (10 mM)

Omit ATP from the ligation reaction in Step 5 if the ligation buffer contains . . . [Full Text of this Article]


METHOD


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