UW-Stout/Heat Shock

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Preliminaries

None 

Materials Yeast genes of each group, Hot Water bath, Water bath tube holders

Wild Type Procedure

1) Set the water bath to 37 degrees celsius and wait for it to heat up 2) Once it is set at the correct temperature, place the yeast samples into it (100-microliter samples) Take samples are 0 minutes, 15, 30, 45, an hour, an hour 15 minutes, and an hour and a half After the hot water bath, pipettee yeast out and place into an assay plate

Preliminaries

None 

Calibration Materials 1) Wild type yeast (we used BY4735) 2) Hot water bath 3) Water bath tube holders 4) Assay plate

Calibration Procedure 1) Set the water bath to 37 degrees Celsius and wait for it to heat up 2) Once it is set at the correct temperature, place the yeast samples into it (100-micro-liter samples) 3) Take samples are 0 minutes, 15, 30, 45, an hour, an hour 15 minutes, and an hour and a half 4) After the hot water bath, pipette yeast out and place into an assay plate

Knockout Materials 1) Knocked out yeast gene sample(s) 2) Hot water bath 3) Water bath tube holders 4) Assay plate

Knockout Procedure Day 1 1) Set the water bath to 37 degrees Celsius and wait for it to heat up 2) Pick a time slot from the original calibration in order to test 3) Put all yeast sample(s) in for specified time and wait 4) After time take yeast sample(s) out and put into the assay plate

Knockout Procedure Day 2 1) Set the water bath to 37 degrees Celsius and wait for it to heat up 2) Pick a time slot from the original calibration in order to test (that is different from day 1) 3) Put all yeast sample(s) in for specified time and wait 4) After time take yeast sample(s) out and put into the assay plate