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Team:Groningen/Labwork/23 July 2013
From 2013.igem.org
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| + | All suspensions were white. Decided to do a miniprep + digestion to see is plasmid was present. | ||
| + | Samples 1 and 6 were chosen, 1 to check if renewed 2012 inoculation was a success and 6 to check if our stock had a the backbone. Upon down spinning the cells (first step of miniprep) a pinkish color was detected in the pellet of cell sample 6. This suggest that the BBa_k818000 backbone was in there after all. Sample 1 showed a less conclusive result. The cells weren't pure white, but also didn't display the same level of pinkness as sample 6. | ||
Revision as of 09:16, 23 July 2013
Mirjam
Run a gel with the PCR products of silk without strep + signal sequence.
The PCR of silk without strep + signal sequence is successful. A new 1.5% agarose gel is made to do gel purification. Did the gel purification.
Run a gel with the purified PCR product of silk strepF-R and silk F-strepR.
The products are present in 2/3 samples for each silk type. These are combined and concentrated. Concentrations of 18.1 and 16.1 ng/ul are obtained. So a restriction digestion can be made.
Made a restriction digestion for the signal sequences.
Sander
Made a restriction digestion for silk strepF-R and silk F-strepR.
Inne
E. coli with the BBa_818000 backbone that had to grow overnight grew.
| Sample | Composition | BBa_k818000 from | growth |
| 1 | 4 mL LB, 8 uL Amp | iGEM 2012 | Yes |
| 2 | 4 mL LB, 8 uL Amp | iGEM 2012 | Yes |
| 3 | 4 mL, LB 8 uL Amp (negative control) | iGEM 2012 | No |
| 4 | 4 mL LB, 8 uL Amp, 4 uL IPTG | iGEM 2012 | Yes |
| 5 | 4 mL LB, 8 uL Amp, 4 uL IPTG | iGEM 2012 | Yes |
| 6 | 4 mL, 8uL Amp | iGEM 2013 | Yes |
| 7 | 4 mL, 8 uL Amp, 8uL IPTG | iGEM 2013 | Yes |
| 8 | 4 mL, 8 uL Amp (negative control) | iGEM 2013 | No |
All suspensions were white. Decided to do a miniprep + digestion to see is plasmid was present. Samples 1 and 6 were chosen, 1 to check if renewed 2012 inoculation was a success and 6 to check if our stock had a the backbone. Upon down spinning the cells (first step of miniprep) a pinkish color was detected in the pellet of cell sample 6. This suggest that the BBa_k818000 backbone was in there after all. Sample 1 showed a less conclusive result. The cells weren't pure white, but also didn't display the same level of pinkness as sample 6.
