plasmids

Joe Boutell boutell at cf.ac.uk
Wed Aug 21 06:40:37 EST 1996


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.




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