Thanks are due Francis Ouellette for keeping this group running!
Quick question: for how long at 4 degrees C are 5-FOA plates still
counterselective for URA3?
Also, how many generations of pregrowth under conditions nonselective
for URA3 are required for a reasonable recovery of transformants if
you are trying to shuffle out a CEN-based plasmid? And can I still use
medium for the pregrowth which is selective for other markers (plasmid
and chromosomal) as long as the medium contains uracil (say in the
concentration found in SD)?
Thanks in advance!
David J. Meyer, Ph.D.
Pioneer Hi-Bred International, Inc.
7300 NW 62nd Ave.
Johnston, IA 50131-1004
Email: meyerdj at phibred.com