I assume you have seen this already
http://www.umanitoba.ca/faculties/medicine/units/human_genetics/gietz/Trafo.
html
Yeast will tend to go petite (lose there mitochondria) when frozen, I am not
sure if this will affect there compatance, but you can select for whole
cells by growing on YEPEG plates.
YEPEG
1. Dissolve 10 gm of succinic acid in 500 ml of distilled H20
2. add 10 gm yeast extract
20 gm bactopeptone
20.8 ml of glycerol
3. adjust pH to 5.5 with 1 M base
4. add 20 gm agar
5. add distilled H20 to 970 ml
6. autoclave
7. when cool add 27.5 ml 95% ETOH
S. Tan wrote in message <808abj$eob at net.bio.net>...
>Hey -
>>We've been doing routine yeast transformations in lab. Recently,
>however, we were unable to transform plasmids into our yeast strains (e.g.
>Hf7c from Clontech). We use fresh medium and new reagents, and have
>tried LiAc method and Bio101 kit. Does anyone know what might be going
>on? Do yeast cells lose "competency" after a long period of storage (at
>-80oC)?
>>Thanks in advance,
>Thomas
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