Download CLONTECH

Yeast Protocols Handbook
CLONTECH Laboratories, Inc.
III. Culturing and Handling Yeast continued
B. Growth Selection for Transformation Markers and Reporter Gene Expression
Most yeast cloning vectors and control plasmids (including those provided in our MATCHMAKER
Systems) carry at least one nutritional marker to allow for selection of yeast transformants plated
on SD minimal medium lacking that specific nutrient. Furthermore, if you are cotransforming yeast
with two or more different plasmids bearing different nutritional markers, the plasmids can be
independently selected. Thus, the SD selection medium you choose for plating transformants
depends generally on the purpose of the selection. Specific factors to consider in choosing the
appropriate SD selection medium are:
•
•
•
•
•
the plasmid(s) used and whether you are selecting for one or more plasmids
whether you are selecting for colonies in which two hybrid proteins are interacting
whether—and to what extent—the host strain is leaky for reporter gene expression
whether you want to induce protein expression from the regulated GAL1 promoter
whether you intend to perform in-vivo, agar-plate β-galactosidase assays (for lacZ
reporter expression in the LexA Two-Hybrid System).
Please refer to your system-specific User Manual for further information on choosing the
appropriate SD selection media for particular plasmids, host strains, and applications.
Protocol # PT3024-1
Version # PR7X265
Technical Support
TEL: 650-424-8222 or 800-662-2566(CLON)
FAX: 650-424-1064 or 800-424-1350
page 11