Competence Test for BW27783 cells
- Take out 100uL glycerol stocks of BW27783 and thaw on ice
- Add 2uL of pUC19 to the BW27783 cells and keep on ice for 30min
- Heat shock at 41.7°C for 60sec
- Put on ice for 2min
- Recover cells in 37°C incubator for 2hours
- Spread plate cells on Amp plate
Transformation of Chloramphenicol Construction Plasmid
- Take out 100uL glycerol stocks of DB3.1 from -80 freezer and thaw on ice
- Add 1uL of Chlor Construction Plasmid to DB3.1 cells and keep on ice for 30min
- Heat shock at 41.7 for 60sec
- Put on ice for 2 min
- Recover cells in 37°C incubator for 2 hours
- Spread plate 500uL on Chlor plate
Results of Chloramphenicol Testing
- Plates were taken from the Lagally Lab 37ºC incubator at 10:05 am.
| Chlor 0||LB-only||Yes||Yes
| Chlor 1||LB + 25µg/mL Chloramphenicol||No||No
- Obviously, the chloramphenicol works. Now, I will aliquot Paul's batch into 1 mL aliquots
- Discarded old batch of Chlor-25 into biohazardous waste.
- Leftover stock kept in -20ºC freezer
- To help Heather with transformation of Chloramphenicol construction plasmid, I will spread plate 5 chlor plates with 25 µL of Chlor-25 stock on each plate.