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Virginia Commonwealth/22 July 2009
From 2009.igem.org
Contents |
Wednesday 22 July 2009
Results
Maria and Afton
- 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
Tasks
Maria and Afton
- 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
Maria and Afton
- Dilute overnight cultures
- 1 mL of sample was diluted with 6 mL of LB + Cm
- only seven samples were saved however, most of these parts were recultured and the flow cytometer was not used today, so these cell cultures most likely will not be kept
- Samples from cryogenic stocks were grown up in LB:
- DB 3.1 (2 vials)
- Samples from cryogenic stocks were grown up in LB + Amp:
- pSB4C5
- 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)
