alpha factor arresting in bar1 strains is a fairly common practice so
i think that others must see this effect also. the first time i did
the arrest the cells did not release synchronously and also did not
release until 45-60min after washout (at 60min many cells were small
budded but others we still shmooing). the next time i used a protease,
pepsin, i washed 1x in YPR, 2x pepsin in YPR, 1x in YPR then released
into YPR. still the cells did not release until after 45-60min but at
least this time they were more synchronous but not quite perfect. ive
ordered the pronase E protease that the koshland webpage recommends
but im not sure if the delayed in the release of the cells from G1 is
common for others. can anyone advise here?
****************************************
HR Silver
PhD Candidate
Department of Biochemistry and Molecular Biology
Thomas Jefferson University
233 South 10th Street, BLSB 231
Philadelphia, Pennsylvania 19107
