Difference between revisions of "Raw Experimental Notes June 2016"
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6/13/2016 - Magi, Kristen, Patrik, Sean | ==6/13/2016 - Magi, Kristen, Patrik, Sean== | ||
Experiment: Yeast Revival & Confirmation of Correct Yeast | Experiment: Yeast Revival & Confirmation of Correct Yeast | ||
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* Preparing bleach: 1:10 bleach to tapwater (grab from restroom; not hand-washing water) | * Preparing bleach: 1:10 bleach to tapwater (grab from restroom; not hand-washing water) | ||
06/15/16 Patrik & Magi | ==06/15/16 Patrik & Magi== | ||
Adding leucine at 120mg/L in a 160mL mix which equates to 19.2mg of Leucine- used 23.4mg (accuracy achieved on scale) | Adding leucine at 120mg/L in a 160mL mix which equates to 19.2mg of Leucine- used 23.4mg (accuracy achieved on scale) | ||
In 160mL we added: 23.4mg of Leucine, 3.2g of Agar, 1.15g of SD-Ura-Glucose and 3.2g of Glucose | In 160mL we added: 23.4mg of Leucine, 3.2g of Agar, 1.15g of SD-Ura-Glucose and 3.2g of Glucose | ||
Revision as of 03:17, 16 June 2016
6/13/2016 - Magi, Kristen, Patrik, Sean
Experiment: Yeast Revival & Confirmation of Correct Yeast
Tasks:
- 0. Revive yeast cultures (Patrik did this previously, and confirmed that the baker's yeast (control) was able to produce its own Uracil; however, the revived cell did not appear to exhibit much growth, so additional 500l of yeast strain was added to the tube, and another tube was also made) - Complete
- 1. Prepare agar mixes (done) - Complete
- 2. Pour Plates - Complete
- 3. Streak plates & then check for colony growth - Pending
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)
06/15/16 Patrik & Magi
Adding leucine at 120mg/L in a 160mL mix which equates to 19.2mg of Leucine- used 23.4mg (accuracy achieved on scale)
In 160mL we added: 23.4mg of Leucine, 3.2g of Agar, 1.15g of SD-Ura-Glucose and 3.2g of Glucose