cycloheximide selection in Y2H

Miguel Aguilera maguilera at iib.uam.es
Tue Aug 25 08:47:16 EST 1998


Hello everybody!

Did anybody use the cycloheximide dominant selection marker in vector
pAS2-1 of the Matchmaker-2 kit? Does it work in the first generation or
several passes are needed to recover the colonies that lost this plasmid?

I have no experience with yeast, but I thought this was a good way to
start recovering putative interactor's plasmids, because in one step you
can test for the loose of B-gal activity of the positive colony, and to
get rid of the bait plasmid for the isolation and analysis of the clone.
Is this all correct or did I forget anything?

Thanks for your help,


Miguel


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Miguel Aguilera
Instituto Cajal, CSIC
Madrid - SPAIN
E-mail: maguilera at iib.uam.es
------------------------------------
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Miguel Aguilera Merlo
Laboratorio de Neurogenetica del Desarrollo
Instituto de Neurociencias "Santiago Ramon y Cajal" (C.S.I.C.)
Ave. Dr. Arce, 34   -   28002 MADRID

Tfno: 34-91-5854738
Fax:  34-91-5854754
E-mail: maguilera at iib.uam.es




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