Smash & Grab
ez003450 at othello.ucdavis.edu
ez003450 at othello.ucdavis.edu
Thu Jun 16 14:39:06 EST 1994
I just discovered an easy way to increase the efficiency of E. coli
transformation when smashing and grabbing plasmids from Yeast. After
collecting the supernatant from the Yeast lysis step, precipitate it with
ethanol and resuspend in TE. Then purify the DNA extract using Wizard
miniprep columns from Promega. The purification on the miniprep columns
only takes about 5 minutes and seems to remove inhibitors of E. coli
transformation resulting in a large(>20 fold) increase in transformation.
Using this modification I was able to get E. coli transformants from
extracts that gave me no transformants before purification.
More information about the Yeast
mailing list