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Virginia Commonwealth/14 July 2009
From 2009.igem.org
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Contents |
Tuesday 14 July 2009
Results
Maria and Afton
- All of the promoters showed up on the gel electrophoresis in a mostly straight line.
- Transformation Results
| Name | Plate | Growth Observation |
|---|---|---|
| Control (+) NEB10β | LB | lawn |
| Control (+) NEB10β shocked | LB | lawn |
| Control (-) NEB10β | Cm | no growth |
| J23106, J06702, pSB4C5 ligation | Cm | ~30 small colonies |
- Miniprep Results:
| Part No. | A260 (nm) | A280 (nm) | A260/A280 | Vol. (µL) | Conc. (µg/mL) | Amt (µg) |
|---|---|---|---|---|---|---|
| J23101 | 0.050 | 0.039 | 1.20 | 47 | 75 | 4.70 |
| J23102 | 0.030 | 0.025 | 1.20 | 58 | 45 | 3.48 |
| J23103 | 0.044 | 0.032 | 1.38 | 56 | 66 | 4.93 |
| J23104 | 0.033 | 0.028 | 1.18 | 54 | 49 | 3.56 |
| J23105 | 0.037 | 0.029 | 1.28 | 32 | 56 | 1.78 |
| J23107 | 0.054 | 0.040 | 1.35 | 47 | 81 | 3.81 |
Trentay 13:44, 14 July 2009 (UTC)
- Gel Electrophoresis Results
Kevin and Adam
- Miniprep results:
| Part No. | A260 (nm) | A280 (nm) | A260/A280 | Vol. (µL) | Conc. (µg/mL) | Amt (µg) |
|---|---|---|---|---|---|---|
| E0240 | .033 | .019 | 1.737 | 124 | 66 | 8.184 |
| pSB4C5 | .054 | .036 | 1.5 | 123.5 | 108 | 13.34 |
Bussingkm 15:21, 14 July 2009 (UTC)
Craig and Clay
- The ampicillin plates left overnight again displayed no growth. The electrotransformed parts will be re-plated onto ampicillin-selective plates as well as non-selective plates.
Tasks
Maria and Afton
- Digest promoters: 101, 102, 103, 104
- Possible Ligation of promoters with parts J06702, pSB4C5
Trentay 15:13, 14 July 2009 (UTC)
Kevin and Adam
- perform complete digestion and ligation
- construct biobrick device (pSB4C5, BBa_E0240, BBa_J23100), and (pSB4C5, BBa_E0240, BBa_J23106)
Bussingkm 15:20, 14 July 2009 (UTC)
Craig and Clay
- The electrotransformed parts will be re-plated on ampicillin plates as well as on non-selective plates. Growth on non-selective plates will prove the existence of viable cells.
Wetlab
Maria and Afton
- Spectophotometry of promoters: 101, 102, 103, 104, 105, 107
- Digestion/Ligation of promoters
- Electrophoresis of promoter digestions: 101, 102, 103, 104, 105, 107, (100)
- Electrotransformation of ligated parts
- two Cm plate had an unidentified growth in the refrigerator.
- however, negative controls from the same plate stock show antibiotic is working
- Picked colonies for overnight culture of J23106 w/ J06702 reporter
- Made 3 overnight 5mL cultures of NEB 10 beta cell stock to prepare electrocompetent cells
Trentay 21:01, 14 July 2009 (UTC)
Kevin and Adam
- pSB4C5, BBa_E0240, and BBa_J06702 were frozen in box C-11 of the glycerol stocks
Bussingkm 15:22, 14 July 2009 (UTC)
Craig and Clay
- The electrotransformed parts were again plated on ampicillin-selective plates. They were also plated on non-selective media. One hundred microliters was used for each plate rather than the standard 35 microliters.
