Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot3836

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Genetic Engineering with PCR

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

This method describes how to modify the termini of PCR products by introducing restriction sites and other features. To reduce the chance of contamination with exogenous DNAs, prepare and use a special set of reagents and solutions for PCR only. Bake all glassware for 6 hours at 150°C and autoclave all plasticware.


MATERIALS

recipe 10x Amplification buffer

Include 0.01% (w/v) gelatin in the buffer.

Oligonucleotide primer 1 (10 µM) in TE (pH 8.0)

Oligonucleotide primer 2 (10 µM) in TE (pH 8.0)

Positive control DNA

Restriction endonucleases

Please see Step 4.

Template DNA

The template DNA could be a cloned gene or . . . [Full Text of this Article]


METHOD


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