Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot4177

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Chromatin Immunoprecipitation in Yeast

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

This protocol was adapted from "Chromatin Immunoprecipitation," Techniques and Protocols 18, 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 100 words of the full text of this article appear below.


INTRODUCTION

Chromatin immunoprecipitation is a method for determining whether a particular protein binds to a particular DNA sequence in vivo.


MATERIALS

Reagents

Appropriate yeast cells to be assayed (1 x 109 - 2 x 109 cells per sample)

caution caution Chloroform:isoamyl alcohol (24:1)

Ethanol, 70% and 100%

caution Formaldehyde (37%)

Glycine, 2.5 M in H2O

Glycogen, 10 mg/ml in H2O

Growth medium

recipe IP wash solution

recipe Lithium chloride (4 M)

recipe Lysis/immunoprecipitation (IP) buffer , ice cold

recipe recipe Lysis IP buffer/500 mM NaCl

caution caution caution Phenol:chloroform:isoamyl alcohol (25:24:1)

PCR reagents

Primary antibody

Protease inhibitors:

Bacitracin, 50X stock solution (50 mg/ml)

Benzamidine, 50X stock solution (50 mM)

caution PMSF, 50X stock . . . [Full Text of this Article]

Equipment


METHOD


TROUBLESHOOTING


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