From 2009.igem.org
Wednesday 22 July 2009
Results
- Overnight cultures were successful
- Vials had noticeable amounts of white E. coli at the bottom of the tube.
- Flow Cytometer was repaired
- Samples cannot be tested until protocols for using and maintaining the machine are figured out
- Plated bacteria are still not expressing GFP and RFP
Trentay
Tasks
- Dilute E. coli into fresh LB + Cm in preparation for testing with the flow cytometer
- nothing was tested today with the cytometer
- Grow all BioBrick parts in 5 mL of LB + Cm overnight
- 100 microliters was added to broth
- plastic tubes had tops, so tops were loosened during the incubation period
- cells were out of the -80 degree Celsius freezer for a prolonged amount of time, so viability is lower than usual.
- If nothing grows, BioBrick DNA will be reinserted into fresh NEB 10 beta through electrotransformation.
- Retransform J23100 w/ J06702
- Transform NEB 10 beta and DB 3.1 to check for electrocompetency
- Organize notebook and running draft
Trentay 19:57, 22 July 2009 (UTC)
Wetlab
- Dilute overnight cultures
- 1 mL of sample was diluted with 6 mL of LB + Cm
- Samples from cryogenic stocks were grown up in LB:
- Samples from cryogenic stocks were grown up in LB + Amp:
- Samples from cryogenic stocks were grown up in LB + Cm:
| Promoter | RET | Growth Observation |
|
| J23100
| E0240 | n/a |
|
| J23101
| E0240 | |
|
| J23101
| J06702 | |
|
| J23102
| E0240 | |
|
| J23102
| J06702 | |
|
| J23103
| E0240 | |
|
| J23103
| J06702 | |
|
| J23104
| E0240 | |
|
| J23104
| J06702 | |
|
| J23105
| E0240 | |
|
| J23105
| J06702 | |
|
| J23106
| E0240 | |
|
| J23106
| J06702 | |
|
| J23107
| E0240 | |
|
| J23107
| J06702 | |
|
| J23110
| E0240 | |
|
| J23110
| J06702 | |
|
| J23113
| E0240 | |
|
| J23113
| J06702 | |
|
| J23116
| E0240 | |
|
| J23116
| J06702 | |
|
| R0011
| E0240 | |
|
| R0011
| J06702 | |
|
| R0040
| E0240 | |
|
| R0040
| J06702 | |
|
- Transformation of electrocompetent E. Coli stock and BioBrick with promoter J23100 and RET J06702
| Name | Type | Plate | Purpose |
|
| NEB 10 beta
| seed stock | LB | make more seed stock |
|
| NEB 10 beta
| electro comp. | LB | (+) Control/ make more stock |
|
| NEB 10 beta
| electro comp. | LB | (+) Control shocked |
|
| DB 3.1
| electro comp. | LB | (+) Control shocked |
|
| DB 3.1
| electro comp. | LB | (+) Control/make more stock |
|
| NEB 10 beta
| electro comp. | Cm | (-) Control |
|
| NEB 10 beta
| electro comp. | Cm | J23100 w/ J06702 |
|
Trentay 20:51, 22 July 2009 (UTC)
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