From 2009.igem.org
Thursday 30 July 2009
Results
Maria and Afton
- The overnight culture grew, however, no RFP was expressed (I1352)
- Spectrophotometry measurements
| Part No. | A260 (nm) | A280 (nm) | A260/A280 | Vol. (µL) | Conc. (µg/mL) | Amt (µg)
|
| pSB4C5 | 0.125 | 0.076 | 1.65 | 27 | 250 | 6.75
|
| I1352 | 0.017 | 0.011 | 1.55 | 96 | 34 | 3.26
|
- Gel Electrophoresis Results
from left to right: pSB4C5, I1352, 100bp Ladder, Bioline Hyperladder
Trentay 14:26, 31 July 2009 (UTC)
Tasks
Maria and Afton
- Make Cryogenic Stocks of part I1352 (2 vials)
- Make another overnight culture from the previous culture to see if it will express RFP
- Miniprep I1352
- Take spectrophotometry measurements of I1352 and pSB4C5
- Digest and Ligate I1352 and pSB4C5
- Run a Gel Electrophoresis on pSB4C5 and I1352 using the Hyper Ladder and the 100bp ladder
- Transform parts into NEB 10 beta
Trentay 14:26, 31 July 2009 (UTC)
Wetlab
Maria and Afton
- Cryogenic stocks were made
- Overnight culture of I1352 was grown up in 5mL of LB+AMP
- 2 samples of 1.5mL of I1352 was Miniprepped
- Spectophotometry measurements were taken
- The two samples were combined into one tube before measurements were taken
- pSB4C5 that was miniprepped on July 27, 2009
- Digestion was done
- Only the restriction enzymes EcoRI-HF and Pstl were used to cut the separate the backbone from the promter and RET
- Ligation was done
- for the Ligation of I1352 with pSB4C5, 3 microliters was added of each digestion to the Ligation sample
- for the Ligation of I1352 with itself, 6 microliters of the Digestion was addes to the Ligation sample
- Gel Electrophoresis was run at 120v for 45 min and DNA bands appeared for both samples
- Transformation was done on the two ligated samples
Trentay 15:10, 31 July 2009 (UTC)
|
"
