From 2009.igem.org
Thursday 16 July 2009
Results
Maria and Afton
- Results of Retransformation with pSB4C5 backbone in NEB 10 beta two days after transformation
| Promoter | Reporter | Plate | Growth Observation
|
| J23101
| E0240 | Cm | no growth
|
| J23101
| J06702 | Cm | 21 colonies
|
| J23102
| E0240 | Cm | 3 colonies
|
| J23102
| J06702 | Cm | 8 colonies
|
| J23103
| E0240 | Cm | 2 colonies
|
| J23103
| J06702 | Cm | 23 colonies
|
| J23104
| E0240 | Cm | 9 colonies
|
| J23104
| J06702 | Cm | 12 colonies
|
| J23105
| E0240 | Cm | no growth
|
| J23105
| J06702 | Cm | 14 colonies
|
| J23107
| E0240 | Cm | 3 colonies
|
| J23107
| J06702 | Cm | 3 colonies
|
- No growth was seen in 5mL culture taken from NEB 10 beta cell freezer stock
- Vial may have thawed out too much after cells were taken out for the electrocompetency procedure
- Spectrophotometry measurements after Miniprep
| Part No. | A260 (nm) | A280 (nm) | A260/A280 | Vol. (µL) | Conc. (µg/mL) | Amt (µg)
|
| J23110 | 0.050 | 0.038 | 1.32 | 48 | 75 | 3.60
|
| J23113 | 0.041 | 0.030 | 1.37 | 44 | 21 | 0.92
|
| J23116 | 0.029 | 0.023 | 1.26 | 44 | 43 | 1.91
|
| R0011 | 0.022 | 0.020 | 1.10 | 41 | 33 | 1.35
|
| R0040 | 0.024 | 0.021 | 1.14 | 42 | 36 | 1.51
|
| E0240 | 0.056 | 0.036 | 1.56 | 109 | 84 | 9.16
|
| pSB4C5 | 0.116 | 0.066 | 1.76 | 174 | 22 | 3.83
|
| J06702 | 0.042 | 0.024 | 1.75 | 63 | 130 | 8.19
|
- Gel Electrophoresis Results
100bp Ladder, Digestions on the Gel: J23110, J23113, J23116, R0011, R0040, E0240
Trentay 14:31, 16 July 2009 (UTC)
Craig and Clay
- No growth was seen on either the kanamycin or chloramphenicol plates.
- The sequence for the LIM1 gene was verified to be optimized for E. coli.
Tasks
Maria and Afton
- Spectrophotometry
- Digest: E0240,J06702, pSB4C5, 110, 113, 116, 0011, 0040
- Ligate promoters with backbone pSB4C5 and reporters E0240, J06702
- Run an electrophoresis test on promoters and reporter Eo240
- Pick colonies of promoters from transformation 100-107 (excluding 106)
- Transformation of NEB 10 beta to make sure new stock is electrocompetent
Trentay 20:41, 16 July 2009 (UTC)
Craig and Clay
- The registry will be contacted to verify the correct resistence for the limonene synthesis related parts. New DNA will also be requested.
Wetlab
Maria and Afton
- Digest: E0240,J06702, pSB4C5, 110, 113, 116, 0011, 0040
- Ligate promoters with backbone pSB4C5 and reporters E0240, J06702
- Run an electrophoresis test on promoters and reporter Eo240
Trentay 20:41, 16 July 2009 (UTC)
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