I am working in a Ts Strain of S. cerevisiae in which the permissive temp is 25c and the non-permissive temp is 36c. I am able to see exponential growth diluting log phase cells once. These cells will grow exponentially again, but when I try to dilute them a second time, they stop growing. The one thing I can think of is a low innoculum causing this as the first dilution is from OD600 of .6 to .3, while the second dilution is from .6 to .03.
Can anyone explain what is going on and what I can do to keep cells growing at 36c?
Yeast Will Not Passage
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