Difference between revisions of "Raw Experimental Notes June 2016"
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6/13/2016 - Maggie, Kristen, Patrik | 6/13/2016 - Maggie, Kristen, Patrik | ||
Experiment: Yeast Revival & Confirmation of Correct Yeast | |||
Tasks: | |||
0. Revive yeast cultures (Patrik did this previously) | |||
1. Prepare agar mixes | |||
2. Pour Plates | |||
ToDo(s): | |||
Immediate: Confirm that the yeast cultures exhibit the correct behavior (ability to grow in 5-FOA and not in SD-URA) | |||
Next Steps: Make new glycerol stock | |||
Later (Next Week): transformation (assuming everything goes well) | |||
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* YPD = 50 g/L; SD- = 7.47 g/L; (need to add 20 g/L of glucose); 5-FOA: 0.5% see prep calculations here: https://docs.google.com/spreadsheets/d/1SRrFP4an20c3uaTYZRlUEeXM6md2O3ES7In1e4Jjqnk/edit#gid=0 | * YPD = 50 g/L; SD- = 7.47 g/L; (need to add 20 g/L of glucose); 5-FOA: 0.5% see prep calculations here: https://docs.google.com/spreadsheets/d/1SRrFP4an20c3uaTYZRlUEeXM6md2O3ES7In1e4Jjqnk/edit#gid=0 | ||
* Last night Patrik started some cultures | * Last night Patrik started some cultures: 1) control 2) revived culture (from the glycerol stock): should be able to grow in presence of 5-FOA but not in SD-URA | ||
* Autoclave settings: 121 C (setting at 21) | * Autoclave settings: 121 C (setting at 21) | ||
Revision as of 04:39, 14 June 2016
6/13/2016 - Maggie, Kristen, Patrik
Experiment: Yeast Revival & Confirmation of Correct Yeast
Tasks: 0. Revive yeast cultures (Patrik did this previously) 1. Prepare agar mixes 2. Pour Plates
ToDo(s): Immediate: Confirm that the yeast cultures exhibit the correct behavior (ability to grow in 5-FOA and not in SD-URA) Next Steps: Make new glycerol stock Later (Next Week): transformation (assuming everything goes well)
- Found previous prepared plates (large & labeled RVC)
- Check for contamination: 4 -URA + Glucose (colorless), 5 (mixture of YPD + YPD + FOA):> decided not to use these and to repaired
- Pouring medias: YPD, SD (-Ura -Glu), 5-FOA; decided to pour whole pack (25), so 8 plates of each
- Volume of plate: 700 ml per plate; decide to make 20 ml of working media (so 20 x 8 plates = 160 ml)
- YPD = 50 g/L; SD- = 7.47 g/L; (need to add 20 g/L of glucose); 5-FOA: 0.5% see prep calculations here: https://docs.google.com/spreadsheets/d/1SRrFP4an20c3uaTYZRlUEeXM6md2O3ES7In1e4Jjqnk/edit#gid=0
- Last night Patrik started some cultures: 1) control 2) revived culture (from the glycerol stock): should be able to grow in presence of 5-FOA but not in SD-URA
- Autoclave settings: 121 C (setting at 21)
Practices:
- Plates should be stored upside to avoid collecting contamination on lids)
- Contaminated plate submerged in bleach water
- Preparing bleach: 1:10 bleach to tapwater (grab from restroom; not hand-washing water)