Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot4169

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PCR-Mediated Gene Disruption: One-Step Method

David C. Amberg, Daniel J. Burke, and Jeffrey N. Strathern

This protocol was adapted from "PCR-Mediated Gene Disruption" Techniques and Protocols 14, in Methods in Yeast Genetics, 2005 edition, by David C. Amberg, Daniel J. Burke, and Jeffrey N. Strathern. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2005.

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


INTRODUCTION

DNA prepared by PCR-mediated gene disruption can be used to transform yeast in gene replacement experiments. This protocol uses two primers, tailed with approximately 50 nucleotides homologous to the gene of interest, that target insertion of the PCR product to that locus. Each primer ends with a universal sequence that is designed to amplify various selectable markers from plasmid templates. Gene . . . [Full Text of this Article]


MATERIALS

Reagents

Equipment


METHOD


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