Yeast Protocols Handbook

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Yeast Protocols Handbook FOR RESEARCH USE ONLYPT3024-1 (PR742227)Published 11 February 2008Yeast Protocols Handbook Clontech Laboratories, Inc. Protocol No. PT3024-1 2 Version No. PR742227 I. Introduction 4 II. Introduction to Yeast Promoters 5 III. Culturing and Handling Yeast 10 IV. Preparation of Yeast Protein Extracts 12 A. General Information 12 B. Preparation of Yeast Cultures for Protein Extraction 12 C. Preparation of Protein Extracts: Urea/SDS Method 13 D. Preparation of Protein Extracts: TCA Method 15 E. Troubleshooting 17 V. Yeast Transformation Procedures 18 A. General Information 18 B. Reagents and Materials Required 19 C. Tips for a Successful Transformation 20 D. Integrating Plasmids into the Yeast Genome 20 E. Small-scale LiAc Yeast Transformation Procedure 20 F. Troubleshooting Yeast Transformation 22 VI. - and -Galactosidase Assays 23 A. General Information 23 B. In vivo Plate Assay Using X-gal in the Medium 25 C.

V2010-1); Molecular Biology and Genetic Engineering of Yeasts, edited by Heslot & Gaillardin (1992); Stargell & Struhl (1996); and Pringle et al. (1997; No. V2365-1). UAS and TATA regions are basic building blocks of yeast promoters The initiation of gene transcription in yeast, as in other organisms, is achieved by several

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