Rapamycin Sensitivity Assay of Yeast - Revision history

Davebrid: added notes about dilutions and incubations

2009-10-14T18:21:04Z

added notes about dilutions and incubations

← Older revision		Revision as of 18:21, 14 October 2009
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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

Davebrid at 15:05, 13 October 2009

2009-10-13T15:05:03Z

← Older revision		Revision as of 15:05, 13 October 2009
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	==Materials==		==Materials==
	*YPD Media		*YPD Media. See [[YPD Media and Agar]]
	*YPD Plates with either DMSO or 2-200 nM Rapamycin		*YPD Plates with either DMSO or 2-200 nM Rapamycin. See [[YPD Media and Agar]]
	*Yeast strains, streaked out on appropriate plate (< 1 month old)		*Yeast strains, streaked out on appropriate plate (< 1 month old)

Davebrid: Created page with '==Materials== YPD Media YPD Plates with either DMSO or 2-200 nM Rapamycin *Yeast strains, streaked out on appropriate plate (< 1 month old) ==Protocol== #Innoculate 3 mL of YP...'

2009-10-13T13:28:30Z

Created page with '==Materials== *YPD Media *YPD Plates with either DMSO or 2-200 nM Rapamycin *Yeast strains, streaked out on appropriate plate (< 1 month old) ==Protocol== #Innoculate 3 mL of YP...'

New page

==Materials==
*YPD Media
*YPD Plates with either DMSO or 2-200 nM Rapamycin
*Yeast strains, streaked out on appropriate plate (< 1 month old)

==Protocol==
#Innoculate 3 mL of YPD in a small erlenmeyer flask with one colony of yeast
#Grow for ~8h at 24C
#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.
#For each flask innoculate several dilutions

Rapamycin Sensitivity Assay of Yeast - Revision history

Davebrid: added notes about dilutions and incubations

Davebrid at 15:05, 13 October 2009

Davebrid: Created page with '==Materials== *YPD Media *YPD Plates with either DMSO or 2-200 nM Rapamycin *Yeast strains, streaked out on appropriate plate (< 1 month old) ==Protocol== #Innoculate 3 mL of YP...'

Davebrid: Created page with '==Materials== YPD Media YPD Plates with either DMSO or 2-200 nM Rapamycin *Yeast strains, streaked out on appropriate plate (< 1 month old) ==Protocol== #Innoculate 3 mL of YP...'