Yeast One Hybrid Screens - (Jan/09/2008 )

I am just starting up a new project looking at transcriptional regulation in zebrafish and will be using yeast one hybrid screening kit to look at this (Clontech Product number 630304). I've never done any yeast work so if anyone has used this kit or has any general tips or good links then your help would be greatly appreciated.

I am already having some problems with media, specifically finding differing recipies for SD medium ?

cheers in advance for any help

Don't know much about doing yeast ONE hybrid, but for yeast two hybrid, this is a great site. I imagine that some of the techniques would be the same.

http://home.cc.umanitoba.ca/~gietz/

I've done yeast one-hybrid screens but without kit, so I suppose there may be differences...

About the minimal medium (SM) doesn't your kit have a recipe for that?
Try to find a protocol where they use the same yeast strain as you do. Using the medium they use will probably be ok.

Hi,

thanks for the help maybe you can help me with this one as well. I have streaked my yeast on a YPDA plate and have very few small colonies (after 4 days), from the manual I was expecting clonies to be 2-3mm. I autoclaved for 15min @ 121C as instructed by the manual,pH before autoclaving was 6.45, some protocols are saying to just autoclave water/agar and then add theYPDA solution by filter sterilization, which is correct/most appropriate? Also after a cople of days the plates were very thin, I poured to approx 1cm but they are now very thin ?

Any hints or tips would be appreciated 1

cheers in advance

After 1 day in the incubator, put the plates into the plastic bags they came in (before pouring) to reduce evaporation.

thanks for the advice guys, am trying as i speak

thanks for the advice, i have what look like nice healthy colonies now