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Team:UNIPV-Pavia/Notebook/Week6
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*We decided to avoid Antarctic Phosphatase treatment in next ligations to save time. | *We decided to avoid Antarctic Phosphatase treatment in next ligations to save time. | ||
| + | |||
| + | *We also decided to cut up to 1 µg of vector, to reduce background noise. | ||
*We choose 4th and 7th colonies to grow 9 ml cultures overnight. | *We choose 4th and 7th colonies to grow 9 ml cultures overnight. | ||
| - | *We also infected 9 ml LB + Amp | + | *We also infected 9 ml LB + Amp with 30 µl of: |
| + | {| | ||
| + | |BBa_E0240 | ||
| + | |BBa_C0078 | ||
| + | |BBa_B0030 | ||
| + | |BBa_J23100 | ||
| + | |} | ||
| + | *glycerol stocks. We incubated all the 6 cultures at 37°C, 220 rpm. | ||
*We transformed the remaining ligations: | *We transformed the remaining ligations: | ||
| Line 52: | Line 61: | ||
**'''BBa_B0030'''-BBa_E1010 | **'''BBa_B0030'''-BBa_E1010 | ||
**'''BBa_B0030'''-BBa_C0051 | **'''BBa_B0030'''-BBa_C0051 | ||
| + | |||
| + | *Wiki updating: Project section. | ||
| + | |||
| + | <br><br> | ||
| + | '''06/24/08''' | ||
| + | <br> | ||
| + | *All the 3 ligation plates showed carpets. We decided to streak plates to produce single colonies plates. We incubated the 3 single colonies plates at 37°C overnight. | ||
| + | |||
| + | *Glycerol stocks for: | ||
| + | {| | ||
| + | |BBa_E0240 | ||
| + | |BBa_C0078 | ||
| + | |'''BBa_B0030'''-BBa_C0061(4) | ||
| + | |- | ||
| + | |BBa_J23100 | ||
| + | |BBa_B0030 | ||
| + | |'''BBa_B0030'''-BBa_C0061(7) | ||
| + | |} | ||
Revision as of 22:28, 6 July 2008
 Home
|  The Team
|  The Project
|  Biological Safety
|  Parts Submitted to the Registry
|
|---|---|---|---|---|
 Dry Lab
|  Wet Lab
|  Modeling
|  Protocols
|  Activity Notebook
|
Notebook
| Week 1 | Week 2 | Week 3 | Week 4 | Week 5 | Week 6 | Week 7 |
|---|
Week 5: 06/23/08 - 06/20/08
06/23/08
- Colony PCR for BBa_B0030-BBa_C0061 and BBa_B0030 (no Ant.Phosph.)-BBa_C0061: 10 colonies for every plate.
- We ran PCR results: some colonies showed both ligated plasmid (heavier band) and false positive plasmid, because it was difficult to pick up single colonies from bacteria carpet. Ligation with Antarctic Phosphatase showed non-pure colonies and 3 false positives. Ligation without Antarctic Phosphatase showed 3 non-pure colonies and 7 pure colonies.
- We decided to avoid Antarctic Phosphatase treatment in next ligations to save time.
- We also decided to cut up to 1 µg of vector, to reduce background noise.
- We choose 4th and 7th colonies to grow 9 ml cultures overnight.
- We also infected 9 ml LB + Amp with 30 µl of:
| BBa_E0240 | BBa_C0078 | BBa_B0030 | BBa_J23100 |
- glycerol stocks. We incubated all the 6 cultures at 37°C, 220 rpm.
- We transformed the remaining ligations:
- BBa_B0030-BBa_E0040
- BBa_B0030-BBa_E1010
- BBa_B0030-BBa_C0051
- Wiki updating: Project section.
06/24/08
- All the 3 ligation plates showed carpets. We decided to streak plates to produce single colonies plates. We incubated the 3 single colonies plates at 37°C overnight.
- Glycerol stocks for:
| BBa_E0240 | BBa_C0078 | BBa_B0030-BBa_C0061(4) |
| BBa_J23100 | BBa_B0030 | BBa_B0030-BBa_C0061(7) |
