Raw Experimental Notes June 2016

6/13/2016 - Maggie, 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 was able to produce its only Uracil) 1. Prepare agar mixes (done) 2. Pour Plates 3. Streak plates & then check for colony growth

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)