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Yeast Protocol Handbook

Yeast Protocol Handbook

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Published by Uday Torres Lamus

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Published by: Uday Torres Lamus on Feb 17, 2010
Copyright:Attribution Non-commercial


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The Yeast Protocols Handbook provides background information and general yeast protocols that
complement our system-specifc User Manuals. The protocols in this Handbook have been optimized
with our yeast-based Matchmaker™ Two-Hybrid and One-Hybrid Systems, and Matchmaker
Libraries. The Yeast Protocols Handbook is especially useful for researchers who wish to use
yeast as a vehicle for their molecular biology experiments, but have little or no prior experience
working with yeast. For novice and experienced users alike, the Yeast Protocols Handbook will
help you obtain the best possible results with your Matchmaker and other yeast-related products
from Clontech.

This Handbook includes:

• detailed information on culturing and handling yeast

• information on the yeast promoters used in the Matchmaker Systems

• two protocols for preparing protein extracts from yeast

• quantitative and qualitative β-galactosidase assays (for use with lacZ yeast reporter


• a simple, optimized protocol for isolating plasmids from yeast

• PCR amplifcation and yeast colony hybridization protocols for the rapid analysis of positive

clones obtained in a library screening

• a small-scale, lithium acetate yeast transformation protocol

• additional protocols for working with certain yeast plasmids and host strains

The special application of yeast transformation for one- and two-hybrid library screening is covered
in detail in each product-specifc User Manual. The special application of yeast mating for library
screening is covered in the Pretransformed Matchmaker Libraries User Manual.

About our yeast-based products

The Matchmaker GAL4 Two-Hybrid Systems (Cat No. K1604-1, K1605-1, 630303) and LexA Two-
Hybrid System (Cat No. K1609-1) are complete kits for identifying and investigating protein-protein
interactions in vivo using the yeast two-hybrid assay. The Matchmaker One-Hybrid System (Cat
No. K1603-1) provides the basic tools for identifying novel proteins in vivo that bind to a target
DNA sequence such as a cis-acting regulatory element. Matchmaker Two-Hybrid Systems are
compatible with our pBridge Three-Hybrid Vector (Cat No. 630404) for the investigation of tertiary
protein complexes. The Matchmaker Libraries are constructed in vectors that express inserts as
fusions to a transcriptional activation domain, and are thus a convenient resource for researchers
wishing to screen a library using the one- or two-hybrid assays. Pretransformed Matchmaker
Libraries provide an even greater level of convenience for those wishing to perform a two-hybrid
library screening without using large- or library-scale yeast transformations.

Clontech offers an extensive line of kits and reagents that support and complement the
Matchmaker Systems and Libraries. The YeastmakerTM

Yeast Transformation Kit (Cat No. 630439)
includes all the necessary reagents and protocols for effcient transformation using the lithium
acetate method. Also available from Clontech: a selection of GAL4 DNA-binding domain (DNA-BD)
and activation domain (AD) hybrid cloning vectors; the pGilda Vector for use with LexA-based
two-hybrid systems; monoclonal antibodies and sequencing primers; and yeast media, including
Minimal SD Base and many different formulations of Dropout (DO) Supplement. Finally, the pHA-
CMV and pMyc-CMV Vector Set (Cat No. 631604) can be used to confrm protein interactions in
mammalian cells.

For ordering information on these products, please see Chapter XI of this Handbook or the
Clontech Catalog.

Yeast Protocols Handbook

Protocol No. PT3024-1


Clontech Laboratories, Inc.

Version No. PR973283


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