Difference between revisions of "UW-Stout/Formamide SP22"
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===Caution=== | ===Caution=== | ||
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4. In a sterile environment, pipet the liquid (50ul) from the PCR tube into the well cell as well as 50ul of the wild yeast cells | 4. In a sterile environment, pipet the liquid (50ul) from the PCR tube into the well cell as well as 50ul of the wild yeast cells | ||
5. Incubate assess tray at 30 degrees Celsius for --- hours and record data hi | 5. Incubate assess tray at 30 degrees Celsius for --- hours and record data hi | ||
| + | |||
| + | ===Results=== | ||
| + | |||
| + | |||
| + | |||
| + | ==Knockout Yeast Cell Gene== | ||
| + | |||
| + | ===Materials=== | ||
| + | *Neat formamide, | ||
| + | *Sterile water | ||
| + | *0.2ml flat cap PCR tube 10 | ||
| + | *Well plate | ||
| + | *Knock out yeast cells | ||
| + | |||
| + | ===Procedure=== | ||
| + | 1. Grab neat formamide and sterile water | ||
| + | 2. Label PCR tubes 1-9 | ||
| + | 3. Pipet approximately --- formamide into a PCR tube, can put excess in | ||
| + | 4. Pipet 3.0ul of formamide into each of the numbered PCR tubes | ||
| + | 5. Pipet 47.0ul of sterile water into each of the numbered PCR tubes | ||
| + | 6. In a sterile environment, pipet 50ul of liquid from the PCR tubes into well cells. Pipet 50ul of each knockout strain into the approperiate well cells | ||
| + | 7. Incubate assess tray at 30 degrees Celsius for --- hours and record data | ||
| + | |||
| + | ===Results=== | ||
Revision as of 20:08, 7 May 2022
==Wild yeast cell== hi
Contents
Caution
Formamide can cause hi
Materials
- Neat Formamide, 33ul
- Sterile water, 567ul
- 0.2ml flat cap PCR tube, 13
- Well plate
- Wild yeast cells, 600ul
Formamide Concentrations
- 0.5ul formamide + 49.5ul sterile water
- 1.0ul formamide + 49.0ul sterile water
- 1.5ul formamide + 48.5ul sterile water
- 2.0ul formamide + 48.0ul sterile water
- 2.5ul formamide + 47.5ul sterile water
- 3.0ul formamide + 47.0ul sterile water
- 3.5ul formamide + 46.5ul sterile water
- 4.0ul formamide + 46.0ul sterile water
- 4.5ul formamide + 45.5ul sterile water
- 5.0ul formamide + 45.0ul sterile water
- 5.5ul formamide + 44.5ul sterile water
- 0.0ul formamide + 50.0ul sterile water
Procedure
1. Grab neat formamide and sterile water 2. Label 12 PCR tubes 1-12 and the 13th one place at least 33ul of neat formamide inside, can put excess in 3. Pipet appropriate amount of formamide and sterile water into the labeled PCR tubes according to number 4. In a sterile environment, pipet the liquid (50ul) from the PCR tube into the well cell as well as 50ul of the wild yeast cells 5. Incubate assess tray at 30 degrees Celsius for --- hours and record data hi
Results
Knockout Yeast Cell Gene
Materials
- Neat formamide,
- Sterile water
- 0.2ml flat cap PCR tube 10
- Well plate
- Knock out yeast cells
Procedure
1. Grab neat formamide and sterile water 2. Label PCR tubes 1-9 3. Pipet approximately --- formamide into a PCR tube, can put excess in 4. Pipet 3.0ul of formamide into each of the numbered PCR tubes 5. Pipet 47.0ul of sterile water into each of the numbered PCR tubes 6. In a sterile environment, pipet 50ul of liquid from the PCR tubes into well cells. Pipet 50ul of each knockout strain into the approperiate well cells 7. Incubate assess tray at 30 degrees Celsius for --- hours and record data
