Difference between revisions of "UW-Stout/Heat Shock FA21"
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UW-Stout/Heat Shock SP21 - SGD-Wiki (yeastgenome.org) | UW-Stout/Heat Shock SP21 - SGD-Wiki (yeastgenome.org) | ||
| − | ---- | + | ----Media Creation Procedure |
| − | |||
| − | Media Creation Procedure | ||
Measure out 10g of Peptide and put into 500ml jar. | Measure out 10g of Peptide and put into 500ml jar. | ||
| Line 33: | Line 31: | ||
Measure out 5g of Yeast Extract and put into 500ml jar. | Measure out 5g of Yeast Extract and put into 500ml jar. | ||
| − | Measure out 10g of | + | Measure out 10g of Agarose and put into 500ml jar. |
Then add 475ml of H2O into the 500ml jar. | Then add 475ml of H2O into the 500ml jar. | ||
| − | Afterwards bring the jar to a | + | Afterwards bring the jar to a laboratory steam sterilizer and put it in for 30 minutes. |
finally pour the media into plates up to half way. | finally pour the media into plates up to half way. | ||
| − | + | ----Heat shock Procedure | |
Calibration Experiment Procedure | Calibration Experiment Procedure | ||
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Afterwards remove all of the tubes from the thermocycler. | Afterwards remove all of the tubes from the thermocycler. | ||
| − | Dilution Procedure | + | ----Dilution Procedure |
| − | + | In a 96 channel well put in your six yeast strains then put them into the left most column desending from A to F. | |
Then in the rest of the rows add 90 ul of H2O into A-F. | Then in the rest of the rows add 90 ul of H2O into A-F. | ||
| Line 69: | Line 67: | ||
After adding the water take 10 ul of the strains add them to the next column and mix. | After adding the water take 10 ul of the strains add them to the next column and mix. | ||
| − | Plating Procedure | + | ----Plating Procedure |
Label each petri dish accordingly, first plate "A,B,C" and the second plate "D,E,F". | Label each petri dish accordingly, first plate "A,B,C" and the second plate "D,E,F". | ||
Revision as of 12:58, 9 December 2021
Materials----
Thermocycler
96 channel well
Wild-type yeast cells
PCR test tubes
P20 and P200 micro pipetters
Multichannel pipette
Media and Plates (all in 475ml H2O)
500 ul of YPD
10g Peptide
5g Yeast Extract
10g Agarose
UW-Stout/Heat Shock SP21 - SGD-Wiki (yeastgenome.org)
Media Creation Procedure
Measure out 10g of Peptide and put into 500ml jar.
Measure out 5g of Yeast Extract and put into 500ml jar.
Measure out 10g of Agarose and put into 500ml jar.
Then add 475ml of H2O into the 500ml jar.
Afterwards bring the jar to a laboratory steam sterilizer and put it in for 30 minutes.
finally pour the media into plates up to half way.
Heat shock Procedure
Calibration Experiment Procedure
Set thermocycler to 48°C sitting temperature.
Vortex the yeast culture briefly to resuspend the yeast cells.
Label 6 pcr tubes A through F.
Micropipette 90 ul of water into the pcr tubes.
Micropipette 10 ul of each strain
Label each tube “A, B, C, D, E, F”.
Insert all 6 of the tubes into the thermocycler for exactly 42 minutes at 48°C.
Afterwards remove all of the tubes from the thermocycler.
Dilution Procedure
In a 96 channel well put in your six yeast strains then put them into the left most column desending from A to F.
Then in the rest of the rows add 90 ul of H2O into A-F.
After adding the water take 10 ul of the strains add them to the next column and mix.
Plating Procedure
Label each petri dish accordingly, first plate "A,B,C" and the second plate "D,E,F".
Micropipette 5 ul of the strains onto the coordinated petri dish using the .
Place all the petri dishes in the incubator (30°C).
Let sit for 48 hours.
Take out and analyze the growth.
Repeat for 3 trials.
