In article <pablo-061094155931 at 22.214.171.124>, pablo at bioc.rice.edu (Paul
> We have been trying to use alpha factor to synchronize S.cerevisiae, but
> the results have not been satisfactory. We are using bar1 strains and
> 1ug/L alpha factor in mid-log phase cells for 2.5 hours at 24C. The alpha
> factor is filtered out, and the cells are then washed in 2 volumes of YPD
> and resuspended in the orginial volume of YPD and grown up at 24C. From
> the bud index taken at
> various times after release, the cells do not appear to be as well
> synchronized as they should be.
>> Any suggestions/how do you do your alpha factor synchronizations?
We would suggest the following modifications:
1. Arrest them for a longer period of time at 24¡C with alpha factor.
Perhaps 3 to 3.5 hrs.
2. Wash the cells much more extensively (at least 4 times) before
releasing them into YPD, and use cold water rather than YPD.
What is the problem anyway? Do you have a large population of budded cells
even before releasing, or a large population that doesn't release? Please
be more specific with your problem....
J. Kroll And S. Tugendreich