McClean:Chemostat Metabolic Cycle: Difference between revisions
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(New page: ==Overview== This is a guide on how to induce the Metabolic Cccle in FY diploid Yeast Strains (First used on 8/5/11-8/14/11) and subsequently sample the culture for various things. To see...) |
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=Overview= | |||
This is a guide on how to induce the Metabolic Cccle in FY diploid Yeast Strains (First used on 8/5/11-8/14/11) and subsequently sample the culture for various things. To see how to set up a chemostat see [[McClean:Basic_Chemostat_Guide | This is a guide on how to induce the Metabolic Cccle in FY diploid Yeast Strains (First used on 8/5/11-8/14/11) and subsequently sample the culture for various things. To see how to set up a chemostat see [[McClean:Basic_Chemostat_Guide| McClean Lab Chemostat Guide]]. | ||
=Inducing the Cell Metabolic = | |||
==Relevant Chemostat Profiles== | |||
Profile 0: | |||
RPM - 400 | |||
Affluent Tube - Off | |||
pH - Off | |||
Temp - 30 °C | |||
DO - minimal | |||
Profile 1: | |||
RPM - 400 | |||
Affluent Tube - On | |||
pH - Off | |||
Temp - 30 °C | |||
DO - minimal | |||
For Dilution Rate .08 hr<sup>-1</sup> : 2/20 | |||
For Dilution Rate .12 hr<sup>-1</sup> : 3/20 | |||
== Inoculation Conditions== | |||
#Fill the vessel with 300 mL of Media. If the Media doesn't automatically fill in the chemostat when the air pressure is turned on and the affluent tube is connected, connect a large tube to the sterile filter of the media carboy and blow some air into it to increase the pressure. Make sure the affluent tube is UNCLIPPED so it can fill. | |||
#Set the profile to 'Profile 0' and then start the program. | |||
#Using 70% ethanol to sterilize, open one of the valves on the top (use a screw driver if ineccessary). Quickly add 3 mL of Overnight Culture in YPD of the strain of interest. | |||
#Quickly tighten the screw and leave the culture as is. | |||
#After ~48 hours (24 hours to reach saturation and 24 hours for starvation) turn off the chemostat program. Change to Profile 1, and restart the chemostat. | |||
#Wait 10-16 hours | |||
Revision as of 09:41, 14 August 2011
Overview
This is a guide on how to induce the Metabolic Cccle in FY diploid Yeast Strains (First used on 8/5/11-8/14/11) and subsequently sample the culture for various things. To see how to set up a chemostat see McClean Lab Chemostat Guide.
Inducing the Cell Metabolic
Relevant Chemostat Profiles
Profile 0: RPM - 400 Affluent Tube - Off pH - Off Temp - 30 °C DO - minimal
Profile 1: RPM - 400 Affluent Tube - On pH - Off Temp - 30 °C DO - minimal
For Dilution Rate .08 hr-1 : 2/20 For Dilution Rate .12 hr-1 : 3/20
Inoculation Conditions
- Fill the vessel with 300 mL of Media. If the Media doesn't automatically fill in the chemostat when the air pressure is turned on and the affluent tube is connected, connect a large tube to the sterile filter of the media carboy and blow some air into it to increase the pressure. Make sure the affluent tube is UNCLIPPED so it can fill.
- Set the profile to 'Profile 0' and then start the program.
- Using 70% ethanol to sterilize, open one of the valves on the top (use a screw driver if ineccessary). Quickly add 3 mL of Overnight Culture in YPD of the strain of interest.
- Quickly tighten the screw and leave the culture as is.
- After ~48 hours (24 hours to reach saturation and 24 hours for starvation) turn off the chemostat program. Change to Profile 1, and restart the chemostat.
- Wait 10-16 hours
