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 (control) was able to produce its own Uracil; however, the revived cell did not appear to exhibit much growth) 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)