From 2009.igem.org
Wednesday 15 July 2009
Results
- Results of Retransformation with pSB4C5 backbone in NEB 10 beta
| Promoter | Reporter | Resistance/Plate | Growth Observation
|
| J23101
| E0240 | Cm | n/a
|
| J23101
| J06702 | Cm | n/a
|
| J23102
| E0240 | Cm | n/a
|
| J23102
| J06702 | Cm | n/a
|
| J23103
| E0240 | Cm | n/a
|
| J23103
| J06702 | Cm | n/a
|
| J23104
| E0240 | Cm | n/a
|
| J23104
| J06702 | Cm | n/a
|
| J23105
| E0240 | Cm | n/a
|
| J23105
| J06702 | Cm | n/a
|
| J23107
| E0240 | Cm | n/a
|
| J23107
| J06702 | Cm | n/a
|
- Controls with NEB 10 beta
| Control | Resistance | Plate | Growth Observation
|
| Positive (+)
| none | LB | lawn
|
| Electroshocked (+)
| none | LB | lawn
|
| Negative (-)
| none | Cm | no cell growth
|
Trentay 16:59, 15 July 2009 (UTC)
Craig and Clay
- The ampicillin plates again showed no growth. However, the non-selective plates displayed lawn growth.
Tasks
Maria and Afton
- Make more electrocompetent NEB 10 beta
- 500 mL LB autoclaved in 1 L flask
- 1 L LB autoclaved
- 2mL of NEB 10 beta overnight culture was diluted in 4mL of LB broth
- Two bottles of LB Agar were prepared
- Three bottles of LB prepared
- two liquid LB+Amp will be prepared
- one liquid LB+Cm will be prepared
- Let transformed BioBrick parts continue to grow overnight (on plates) before colonies are picked
- Make cell stock of Biobrick part: J23106, J06702, pSB4C5
- 2mL of BioBrick overnight culture was diluted in 4mL of LB broth
- Organize Notebook(s)
Trentay 16:59, 15 July 2009 (UTC)
Craig and Clay
- The cells growing on the non-selective media will be streaked onto plates containing kanamycin and plates containing chloramphenicol.
Wetlab
Maria and Afton
- All LB was prepared and autoclaved
- Overnight culture dilutions were completed
- Electrocompetency of NEB 10 beta was completed
- LB and Sterile water was made
- 10% glycerol stock was made
Trentay 14:13, 16 July 2009 (UTC)
Kevin & Adam
- run gel on digestion from yesterday assembly.
Craig and Clay
- Cells possibly containing the limonene synthesis related parts grown on the plates containing no antibiotics were taken and plated onto kanamycin-selective and chloramphenicol-selective plates.
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