"
Team:UNIPV-Pavia/Notebook/Week5
From 2008.igem.org
 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: 06/16/08 - 06/20/08
06/16/08
- We picked up the only colony in BBa_B0030-BBa_C0078 plate to grow a 9 ml culture of transformed bacteria overnight.
- We also infected 9 ml of LB + suitable antibiotic with 30 µl of:
| BBa_B0030 | BBa_E1010 | BBa_E0040 |
| BBa_C0061 | BBa_C0051 |
- glycerol stocks. We incubated the 9 ml culture overnight at 37°C, 220 rpm.
06/17/08
- Glycerol stocks for:
| BBa_B0030 | BBa_E1010 | BBa_E0040 |
| BBa_C0061 | BBa_C0051 | BBa_B0030-BBa_C0078 |
- Miniprep for all these parts.
- Digestion for:
| BBa_B0030 (S-P) | BBa_E1010 (X-P) | BBa_E0040 (X-P) |
| BBa_C0061 (X-P) | BBa_C0051 (X-P) | BBa_B0030-BBa_C0078 (X-S) |
- Gel run for BBa_B0030-BBa_C0078 to check for insert length: unfortunately, there was not a band where we expected...the only colony was a false positive. We'll try to ligate it in the next days.
- Gel run for:
| BBa_B0030 (S-P) | BBa_E1010 (X-P) | BBa_E0040 (X-P) |
| BBa_C0061 (X-P) | BBa_C0051 (X-P) |
- Gel cut and DNA extraction.
- We put DNA at -20°C. The next day we will perform some ligation reaction in different conditions, looking for the best protocol.
06/18/08
- We gave a lecture about Synthetic Biology and our current work at DIS (Department of Informatics and System Science).
- Antarctic Phosphatase for half of BBa_B0030 (S-P) volume.
- We planned the following ligation experiments:
- Transformation with BBa_B0030(S-P), to check background noise (we will know the amount of not digested vector);
- Transformation with this ligation: BBa_B0030(S-P) after Antarctic Phosphatase treatment and no insert;
- Transformation with this ligation: BBa_B0030(S-P) without Antarctic Phosphatase treatment and no insert;
- Transformation with this ligation: BBa_B0030(S-P) without Antarctic Phosphatase treatment and BBa_C0061 insert;
- Transformation with this ligation: BBa_B0030(S-P) after Antarctic Phosphatase treatment and BBa_C0061 insert;
- 3 Transformation with these ligations: BBa_B0030(S-P) after Antarctic Phosphatase treatment and BBa_C0051, BBa_E1010, BBa_E0040 inserts;
- Antarctic Phosphatase for half of BBa_B0030 (S-P) volume.
- We transformed 60 µl of TOP10 with 1 µl of BBa_B0030 (S-P)
- We plated transformed bacteria and incubated them at 37°C overnight.
- Antarctic Phosphatase for half of BBa_B0030 (S-P) volume.
- Ligation:
