- Put 50mL LB with 1 mL of dH10B (2 500 ml cylinders) in the shaker at 9:23 am. (Vallari Somayaji)
- OD of the flasks were checked at 1:00 p.m. The OD was found to be about 1.4. This is concentration is too high to make competent cells. Because of time constraints, a sample was transferred to culture tube containing 5 ml of LB. New flasks will be made on 6/26 to try again.
- The following parts were picked from the iGEM registry plates:
||K654058 - TesA
||E0240 - GFP Generator
||J06702 - mCherry
||R0040 - TetR Promoter
||R0011 - LacI Promoter
||B0034 - RBS
||B0032 - RBS
- Each part was re-hydrated in 10 μl of water.
- 30 μl of competent cells, donated from the Haynes lab, was added to each DNA sample.
- The 7 samples were left on ice for 30 minutes.
- Heat shocked at 42°C for 35 seconds.
- Placed back on ice for 2 minutes
- 750 μl of LB media was added to each tube
- Tubes were placed in shaking incubator for 1 hour at 32°C
- 350 μl of supernatant was removed from each tube
- Pellets were re-suspended and placed onto LB agar plates with chloramphenicol.