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Virginia Commonwealth/11 August 2009
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==Tuesday 11 August 2009== | ==Tuesday 11 August 2009== | ||
===Results=== | ===Results=== | ||
| + | ''Kevin and Adam'' | ||
*I have cell growth on all plates. Yesterday's work was successful. Colonies number from 2-5 colonies per plate at 4:00 PM. I will wait until later to pick a colony. | *I have cell growth on all plates. Yesterday's work was successful. Colonies number from 2-5 colonies per plate at 4:00 PM. I will wait until later to pick a colony. | ||
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* Transformation was done | * Transformation was done | ||
* Overnight cultures were made | * Overnight cultures were made | ||
| - | * | + | * LB+Cm plates were made |
[[User:Trentay|Trentay]] 20:12, 11 August 2009 (UTC) | [[User:Trentay|Trentay]] 20:12, 11 August 2009 (UTC) | ||
| + | |||
| + | ''Adam and Kevin'' | ||
| + | * The growth on the plates remains slow. No colonies show fluorescence yet, for the sake of better selection I am going to wait until early tomorrow morning to pick colonies. I will cultivate them tomorrow and then store them in the evening. Hopefully we will be able to measure transcription levels Thursday. | ||
| + | -[[User:Bussingkm|Bussingkm]] 21:16, 11 August 2009 (UTC) | ||
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Contents |
Tuesday 11 August 2009
Results
Kevin and Adam
- I have cell growth on all plates. Yesterday's work was successful. Colonies number from 2-5 colonies per plate at 4:00 PM. I will wait until later to pick a colony.
Maria and Afton
- 1kb ladder has not come in yet so, digests and minipreps will be run on gels when it comes in
- Plated colonies from Friday (8/7/09) transformations are strongly espressing RFP
- It might be better to pick colonies 3 to 4 days after transformation when colonies are strongly espressing RFP, instead of when the e. coli are pink the day after transformation
- Spectrophotometry Data
| Part No. | A260 (nm) | A280 (nm) | A260/A280 | Vol. (µL) | Conc. (µg/mL) | Amt (µg) |
|---|---|---|---|---|---|---|
| pSB1C3 p1010 | 0.012 | 0.008 | 1.5 | 54 | 24 | 1.3 |
| J23100 | 0.029 | 0.023 | 1.26 | 35 | 58 | 2.03 |
| J23104 | 0.030 | 0.026 | 1.15 | 38 | 60 | 2.28 |
| J23102 | 0.022 | 0.019 | 1.16 | 40 | 44 | 1.76 |
| J23103 | 0.021 | 0.015 | 1.40 | 45 | 42 | 1.89 |
| J06702 | 0.029 | 0.018 | 1.61 | 117 | 58 | 6.79 |
Trentay 19:17, 11 August 2009 (UTC)
Tasks
Maria and Afton
- Take Spectrophotometry measurements of pSB1C3 p1010, J06702, J23100, J23102, J23103, J23104
- Need to locate J23100 to be digested another day
- Digest above parts
- Ligate J06702 and pSB1C3 with the following promoters: 100, 102, 103, 104
- Transform parts into NEB10β
- Update Wiki
- Make overnight cultures of I1352/pSB1C3
- I1352/I1352 will not be kept because it never expressed RFP and there are plenty of stock of I1352 with Amp resistance
- Make overnight cultures of pSB1C3 p1010 from cryogenic storage to Miniprep tomorrow
- Make more LB+Cm plates
Trentay 14:43, 11 August 2009 (UTC)
Wetlab
Maria and Afton
- Spectrophotometry was done
- Digest was done
- Ligation was done
- Transformation was done
- Overnight cultures were made
- LB+Cm plates were made
Trentay 20:12, 11 August 2009 (UTC)
Adam and Kevin
- The growth on the plates remains slow. No colonies show fluorescence yet, for the sake of better selection I am going to wait until early tomorrow morning to pick colonies. I will cultivate them tomorrow and then store them in the evening. Hopefully we will be able to measure transcription levels Thursday.
-Bussingkm 21:16, 11 August 2009 (UTC)
