Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4157

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Assay of ß-Galactosidase in Yeast: Assay of Crude Extracts

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

This protocol was adapted from "Assay of ß-Galactosidase in Yeast," Techniques and Protocols 8, 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

There are several methods for the in vitro assay of ß-galactosidase in yeast. In the method described here (Rose and Botstein 1983), a crude extract is prepared, and the activity is normalized to the amount of protein assayed. This method is particularly suitable for comparing cells that are grown under very different conditions or that have different genetic backgrounds.


MATERIALS

Reagents

Appropriate yeast culture

Bovine serum albumin (BSA), 0.1-1 mg/ml in breaking buffer

Bradford reagent (Bio-Rad)

recipe Breaking buffer

Liquid growth medium (with selection as appropriate)

Na2CO3 stock solution, 1 M

ONPG (o-nitrophenyl-ß-D-galactoside) stock solution: 4 . . . [Full Text of this Article]

Equipment


METHOD


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