yeast transformations

Anne Spang spang at vms.biochem.mpg.de
Fri Feb 3 16:59:37 EST 1995


Hi John!
We are currently using a lithium acetate-DMSO method
- grow O/N culture in 2 ml
- spin down in a 2 ml-Eppi
- remove supernatant, add 0.01 ml carrier DNA (= 0.1 mg) and transforming
DNA in  minimal volume, vortex briefly
- add 0.5 ml PEG (40% 3350, 0.05 ml 10xLiOAc, 0.05 ml 10xTE, H2O ad 0.5 ml)
- add 0.051 ml DMSO
- shake 15 min RT
- shock cells for 15 min at 42C
- spin down, wash the pellet with TE, resuspend in TE and plate on
selective plates

Hope this helps
Anne



More information about the Methods mailing list