Difference between revisions of "UW Stout/Nystatin SP22"
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Purpose: Using a stock solution of nystatin, DMF, and sterile water to treat wild yeast cells. The results of this will be used to decide on a concentration level to apply to different yeast strains. | Purpose: Using a stock solution of nystatin, DMF, and sterile water to treat wild yeast cells. The results of this will be used to decide on a concentration level to apply to different yeast strains. | ||
| − | Materials: | + | Materials: |
| − | 1. Nystatin powder | + | 1.Nystatin powder |
| − | 2. Dimethylformamide (DMF) | + | 2.Dimethylformamide (DMF) |
| − | 3. Sterile water | + | 3.Sterile water |
| − | 4. 1.7ml Microcenterfuge tubes | + | 4.1.7ml Microcenterfuge tubes |
| − | 5. Weighing boat | + | 5.Weighing boat |
| − | 6. Corning COSTAR 96-well clear flat-bottom assay plate | + | 6.Corning COSTAR 96-well clear flat-bottom assay plate |
| − | 7. Glycerol stock of BY4735 strain as prepared in steps 1-4 of the basic growth curve protocol | + | 7.Glycerol stock of BY4735 strain as prepared in steps 1-4 of the basic growth curve protocol |
| − | Required Equipment: | + | Required Equipment: |
| − | 1. Vortex | + | 1.Vortex |
| − | 2. Scale | + | 2.Scale |
| − | 3. Molecular Devices SpectraMax Plus 384 Microplate Reader set as outlined in the basic growth curve protocol | + | 3.Molecular Devices SpectraMax Plus 384 Microplate Reader set as outlined in the basic growth curve protocol |
| − | Protocol For Pilot Experiment | + | Protocol For Pilot Experiment |
| − | Steps to make stock solution: | + | Steps to make stock solution: |
| − | + | •weigh out 40mg of Nystatin power | |
| − | + | •Add nystatin to tube | |
| − | + | •Measure 10ml DMF | |
| − | + | •Add DMF to tube | |
| − | + | •Measure sterile water | |
| − | + | •Mix until no powder is present | |
| − | Dilutions for pilot experiment | + | Dilutions for pilot experiment |
| − | Steps to make dilutions: | + | Steps to make dilutions: |
| − | 1. Mix sterile water with the stock solution, use table below for the amount of each to use | + | 1.Mix sterile water with the stock solution, use table below for the amount of each to use |
| − | 2. Repeat until all concentrations are complete | + | 2.Repeat until all concentrations are complete |
| − | 3. Add 50ul of the diluted product with 50ul of wild yeast | + | 3.Add 50ul of the diluted product with 50ul of wild yeast |
| − | Dilution table: | + | Dilution table: |
| − | Desired Concentrations | + | Desired Concentrations |
| − | 1. 0ug/ml 0ug/ul | + | 1. 0ug/ml 0ug/ul |
| − | 2. 0.5ug/ml or 0.0005ug/ul | + | 2. 0.5ug/ml or 0.0005ug/ul |
| − | 3. 0.75ug/ml 0.00075ug/ul | + | 3. 0.75ug/ml 0.00075ug/ul |
| − | 4. 1.0ug/ml or 0.001ug/ul | + | 4. 1.0ug/ml or 0.001ug/ul |
| − | 5. 1.25ug/ml or 0.00125ug/ul | + | 5. 1.25ug/ml or 0.00125ug/ul |
| − | 6. 1.5ug/ml or 0.0015ug/ul | + | 6. 1.5ug/ml or 0.0015ug/ul |
| − | 7. 1.75ug/ml or 0.00175ug/ul | + | 7. 1.75ug/ml or 0.00175ug/ul |
| − | 8. 2.0ug/ml or 0.002ug/ul | + | 8. 2.0ug/ml or 0.002ug/ul |
| − | Trials 1 and 2 protocol | + | Trials 1 and 2 protocol |
| − | Purpose: Create a nystatin treatment that is able to slow down the growth of different yeast strains. | + | Purpose: Create a nystatin treatment that is able to slow down the growth of different yeast strains. |
| − | Materials: | + | Materials: |
| − | 1.Nystatin powder | + | 1.Nystatin powder |
| − | 2.Dimethylformamide (DMF) | + | 2.Dimethylformamide (DMF) |
| − | 3.Sterile water | + | 3.Sterile water |
| − | 4.1.7ml Microcenterfuge tubes | + | 4.1.7ml Microcenterfuge tubes |
| − | 5.Corning COSTAR 96-well clear flat-bottom assay plate | + | 5.Corning COSTAR 96-well clear flat-bottom assay plate |
| − | 6.Glycerol stock of BY4735 strain as prepared in steps 1-4 of the basic growth curve protocol | + | 6.Glycerol stock of BY4735 strain as prepared in steps 1-4 of the basic growth curve protocol |
| − | Required Equipment: | + | Required Equipment: |
| − | 1.Vortex | + | 1.Vortex |
| − | 2. Micropipettes | + | 2. Micropipettes |
| − | Protocol for trials 1 and 2: | + | Protocol for trials 1 and 2: |
| − | 1.Measure out stock solution and place it in 1.7 tube | + | 1.Measure out stock solution and place it in 1.7 tube |
| − | 2.Measure out and add water to the tube | + | 2.Measure out and add water to the tube |
| − | 3.Vortex the mixture | + | 3.Vortex the mixture |
| − | 4.Apply 50ul of product to each yeast strain | + | 4.Apply 50ul of product to each yeast strain |
| − | Dilution: | + | Dilution: |
| − | 1ug/ml | + | 1ug/ml |
Revision as of 10:36, 5 May 2022
Pilot Protocol
Purpose: Using a stock solution of nystatin, DMF, and sterile water to treat wild yeast cells. The results of this will be used to decide on a concentration level to apply to different yeast strains.
Materials:
1.Nystatin powder
2.Dimethylformamide (DMF)
3.Sterile water
4.1.7ml Microcenterfuge tubes
5.Weighing boat
6.Corning COSTAR 96-well clear flat-bottom assay plate
7.Glycerol stock of BY4735 strain as prepared in steps 1-4 of the basic growth curve protocol
Required Equipment:
1.Vortex
2.Scale
3.Molecular Devices SpectraMax Plus 384 Microplate Reader set as outlined in the basic growth curve protocol
Protocol For Pilot Experiment
Steps to make stock solution:
•weigh out 40mg of Nystatin power
•Add nystatin to tube
•Measure 10ml DMF
•Add DMF to tube
•Measure sterile water
•Mix until no powder is present
Dilutions for pilot experiment
Steps to make dilutions:
1.Mix sterile water with the stock solution, use table below for the amount of each to use
2.Repeat until all concentrations are complete
3.Add 50ul of the diluted product with 50ul of wild yeast
Dilution table:
Desired Concentrations
1. 0ug/ml 0ug/ul
2. 0.5ug/ml or 0.0005ug/ul
3. 0.75ug/ml 0.00075ug/ul
4. 1.0ug/ml or 0.001ug/ul
5. 1.25ug/ml or 0.00125ug/ul
6. 1.5ug/ml or 0.0015ug/ul
7. 1.75ug/ml or 0.00175ug/ul
8. 2.0ug/ml or 0.002ug/ul
Trials 1 and 2 protocol Purpose: Create a nystatin treatment that is able to slow down the growth of different yeast strains.
Materials: 1.Nystatin powder 2.Dimethylformamide (DMF) 3.Sterile water 4.1.7ml Microcenterfuge tubes 5.Corning COSTAR 96-well clear flat-bottom assay plate 6.Glycerol stock of BY4735 strain as prepared in steps 1-4 of the basic growth curve protocol
Required Equipment: 1.Vortex 2. Micropipettes
Protocol for trials 1 and 2: 1.Measure out stock solution and place it in 1.7 tube 2.Measure out and add water to the tube 3.Vortex the mixture 4.Apply 50ul of product to each yeast strain
Dilution: 1ug/ml
Trial 1 results:
Trial 2 results:
