Transformation with tRNA carrier
Bernard P. Murray, PhD
bpmurray*STUFFER* at socrates.cgl.ucsf.EDU
Wed Mar 3 09:33:17 EST 1999
In article <7bf4qc$590 at net.bio.net>, "R. Stevens"
<rstevens at u.washington.edu> wrote:
> Hey Yeastfolk,
> I need to do yeast transformations (into S. cerevisiae) using
> carrier that doesn't contain genomic DNA. I am wondering how the
> protocol differs if I use tRNA instead of salmon testes or calf thymus
> DNA.
> I have tried a standard LiOAc/PEG transformation with 10ug tRNA
> carrier, and got almost no colonies (and hundreds of colonies on the calf
> thymus DNA control). What's up?
> Take care,
> Rick
Check out the original papers by the Gietz lab on transformation
and you'll find that tRNA just doesn't work. I can't remember
the results with ssDNA (from M13) but this may be a possible
alternative if it *has* to be DNA. Another possibility is to
use long oligos.
I hope you can find something,
Bernard
--
Bernard P. Murray, PhD
Dept. Cell. Mol. Pharmacol., UCSF, San Francisco, USA
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