scamargo at usp.br (Solange S de Camargo) wrote:
>>Hi folks,
>> Thanks for all that replied my message about geneticin. It really
>worked out. One of the strains was sensitive to 50ug/ml and the other to
>100ug/ml in liquid YPD medium. I think it's ok and I intend to analyze
>them in solid medium before the transformation.
> Now I have another question: a colleague of mine wants to
>do a plasmid extraction directly from yeast without the step that
>involves bacteria transformation. Does anybody know about any method? She
>can't select the gene she is cloning in bacteria because it is letal for
>this organism. If somebody could help us, we'd appreciate a lot.
> Thanks in advance,
> Solange
Hiya!
Have a word with QIAGEN about doing a plasmid prep from yeast. I looked
into it a while back, and one of their reps gave me an experimental
protocol (one they haven't tested or optimized). It involves a few extra
steps at the beginning, but it uses one of their midiprep columns. Not
sure what kind of yield you get out at the end as i never tried it myself
(i need to separate the two different plasmids in my yeast before
analysing further). Anyway, get in touch with them.
Cheerio,
Joe Boutell.
