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Virginia Commonwealth/22 July 2009
From 2009.igem.org
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[[User:Trentay|Trentay]] 19:57, 22 July 2009 (UTC) | [[User:Trentay|Trentay]] 19:57, 22 July 2009 (UTC) | ||
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====Wetlab==== | ====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 + Cm: | * Samples from cryogenic stocks were grown up in LB + Cm: | ||
| + | ** J23100 w/ E0240 (taken from plates) | ||
** J23101 w/ E0240 | ** J23101 w/ E0240 | ||
** J23101 w/ J06702 | ** J23101 w/ J06702 | ||
| Line 45: | Line 46: | ||
** J23116 w/ E0240 | ** J23116 w/ E0240 | ||
** J23116 w/ J06702 | ** J23116 w/ J06702 | ||
| - | ** R0011 w/ E0240 | + | ** R0011 w/ E0240 |
| - | ** R0011 w/ J06702 | + | ** R0011 w/ J06702 |
| - | ** R0040 w/ E0240 | + | ** R0040 w/ E0240 |
| - | ** R0040 w/ J06702 | + | ** R0040 w/ J06702 |
* Samples from cryogenic stocks were grown up in LB: | * Samples from cryogenic stocks were grown up in LB: | ||
** DB 3.1 (2 vials) | ** DB 3.1 (2 vials) | ||
* Samples from cryogenic stocks were grown up in LB + Amp: | * Samples from cryogenic stocks were grown up in LB + Amp: | ||
** pSB4C5 | ** pSB4C5 | ||
| + | * Transformation of electrocompetent E. Coli stock and BioBrick with promoter J23100 and RET J06702 | ||
| + | ** NEB 10 beta (seed stock) plated on LB | ||
| + | ** NEB 10 beta (electro comp.) plated on LB | ||
| + | ** NEB 10 beta (electro comp.) plated on LB (shocked) | ||
| + | ** DB 3.1 (electro comp.) plated on LB (shocked) | ||
| + | ** DB 3.1 (electro comp.) plated on LB | ||
| + | ** NEB 10 beta (electro comp.) plated on Cm | ||
| + | ** NEB 10 beta (electro comp.) J23100 w/ J06702 plated on Cm | ||
Revision as of 20:16, 22 July 2009
Contents |
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
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 + Cm:
- J23100 w/ E0240 (taken from plates)
- J23101 w/ E0240
- J23101 w/ J06702
- J23102 w/ E0240
- J23102 w/ J06702
- J23103 w/ E0240
- J23103 w/ J06702
- J23104 w/ E0240
- J23104 w/ J06702
- J23105 w/ E0240
- J23105 w/ J06702
- J23106 w/ E0240
- J23106 w/ J06702
- J23107 w/ E0240
- J23107 w/ J06702
- J23110 w/ E0240
- J23110 w/ J06702
- J23113 w/ E0240
- J23113 w/ J06702
- J23116 w/ E0240
- J23116 w/ J06702
- R0011 w/ E0240
- R0011 w/ J06702
- R0040 w/ E0240
- R0040 w/ J06702
- 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
- Transformation of electrocompetent E. Coli stock and BioBrick with promoter J23100 and RET J06702
- NEB 10 beta (seed stock) plated on LB
- NEB 10 beta (electro comp.) plated on LB
- NEB 10 beta (electro comp.) plated on LB (shocked)
- DB 3.1 (electro comp.) plated on LB (shocked)
- DB 3.1 (electro comp.) plated on LB
- NEB 10 beta (electro comp.) plated on Cm
- NEB 10 beta (electro comp.) J23100 w/ J06702 plated on Cm
