Week 1
Competent Cells
- 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.
Transforming cells
- The following parts were picked from the iGEM registry plates:
Tube
|
Registry Number
|
1 |
K654058 - TesA
|
2 |
E0240 - GFP Generator
|
3 |
J06702 - mCherry
|
4 |
R0040 - TetR Promoter
|
5 |
R0011 - LacI Promoter
|
6 |
B0034 - RBS
|
7 |
B0032
|
|
- 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.
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