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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