Rapamycin Sensitivity Assay of Yeast: Difference between revisions
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#Check OD-600 | #Check OD-600 | ||
#Estimate how much to dilute yeast so that the next morning the OD is 0.3. For example, if the doubling time is 2.25h, then from 5 PM to 9 AM is 16h or 7.1 doublings, of 137 times the OD. Therefore to get an OD of 0.3, you need to innoculate at 0.3/137 = 0.00219. | #Estimate how much to dilute yeast so that the next morning the OD is 0.3. For example, if the doubling time is 2.25h, then from 5 PM to 9 AM is 16h or 7.1 doublings, of 137 times the OD. Therefore to get an OD of 0.3, you need to innoculate at 0.3/137 = 0.00219. | ||
#For each flask innoculate several dilutions | #For each flask innoculate several dilutions. | ||
#The next morning check OD of overnight cultures. Cultures should be between 0.1 and 0.5 | |||
#Spin down cultures and resuspend to an OD of 3.0 | |||
#Make 10X serial dilutions of cultures and plate 1 uL of each dilution on YPD and YPD/Rapamycin Plates | |||
#Grow plates at either 24 or 30C | |||