plasmid isolation from yeast
rennie at bldghsc.lan1.umanitoba.ca
Thu Sep 19 15:02:48 EST 1996
> Yeah, but what was wanted was a protocol where you don't have to
> transform the stuff back into E.coli. In which case i would direct them
> to their local QIAGEN rep., who a while ago gave me an experimental
> protocol for isolating plasmid DNA from yeast (ie. they haven't optimised
> it fully):
> 4. Spin down spheroblasts 5000rpm for 5 minutes.
> 5. Resuspend in 4 ml QIAGEN buffer P1, incubate 37 C for 15 min.
> 6. Add 2 ml lysis buffer (3% SDS, 60 mM EDTA). Mix gently but thoroughly,
> incubate 20 min 65 C. Volume should be 6 ml.
> 9. Follow QIAGEN Plasmid Midi protocol starting at equilibration step.
> Joe Boutell. (no affiliation to QIAGEN, but a free t-shirt or keg of beer
> wouldn't go amiss!!?).
I just thought that maybe people would want to know the reference for this
"experimental" protocol minus the "Qiagen buffers and columns"
Our lab calls them "Clyde preps" the real reference is found in:
Cryer,D.R. Eccleshal, R. and Marmur, J. (1975) Isolation of yeast DNA. Meth.
Cell. Biol. 12. pp39-44.
Hope this helps D.R. Cryer (sorry Qiagen)
More information about the Yeast