Difference between revisions of "UW Stout/Proline FA19"
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*Corning COSTAR 96-Well Clear Flat-Bottom Assay Plate | *Corning COSTAR 96-Well Clear Flat-Bottom Assay Plate | ||
==Protocol== | ==Protocol== | ||
| − | ===Part 1=== | + | ===Part 1- Making Proline and Ammonium Sulfate Media=== |
#Label two separate 250ml glass media jars with Proline and Ammonium Sulfate | #Label two separate 250ml glass media jars with Proline and Ammonium Sulfate | ||
#In the Proline jar mix the following materials | #In the Proline jar mix the following materials | ||
| Line 32: | Line 32: | ||
#Autoclave at 120 degrees Celsius for 15 minutes | #Autoclave at 120 degrees Celsius for 15 minutes | ||
#Mix 5ml of glucose to both solutions | #Mix 5ml of glucose to both solutions | ||
| + | |||
===Part 2=== | ===Part 2=== | ||
#In biosafety cabinet pipet 1ml of each culture into separate microcentrifuge tubes twice | #In biosafety cabinet pipet 1ml of each culture into separate microcentrifuge tubes twice | ||
Revision as of 17:16, 17 December 2019
Materials
- Histidine
- Leucine
- uracil
- CSM
- YBN
- Glucose
- Praline
- Ammonium Sulfate
- Water
- Corning COSTAR 96-Well Clear Flat-Bottom Assay Plate
Protocol
Part 1- Making Proline and Ammonium Sulfate Media
- Label two separate 250ml glass media jars with Proline and Ammonium Sulfate
- In the Proline jar mix the following materials
- Histidine 100ml
- Leucine 100ml
- Uracil 100ml
- CSM 0.065g
- YBN 0.171g
- Proline 0.1g
- Fill up to 250ml with water
- In the Ammonium Sulfate jar mix the following materials
- Histidine 100ml
- Leucine 100ml
- Uracil 100ml
- CSM 0.065g
- YBN 0.171g
- Ammonium Sulfate 0.5g
- Fill up to 250ml with water
- Mix well
- Autoclave at 120 degrees Celsius for 15 minutes
- Mix 5ml of glucose to both solutions
Part 2
- In biosafety cabinet pipet 1ml of each culture into separate microcentrifuge tubes twice
- Centrifuge tubes of 30 seconds at 5000 RCF
- In biosafety cabinet aspirate supernatant out
- Label tubes with Proline or Ammonium Sulfate
- Add 1ml of either Proline or Ammonium Sulfate
- Vortex Solutions
- Pipet 100ul of each solution in to wells on the microplate
