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Team:UNIPV-Pavia/Notebook/Week11
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Notebook
| Week 1 | Week 2 | Week 3 | Week 4 | Week 5 | Week 6 | Week 7 |
|---|---|---|---|---|---|---|
| Week 8 | Week 9 | Week 10 | Week 11 | Week 12 | Week 13 | Week 14 |
Week 11: 07/28/08 - 08/1/08
07/28/08
- Colony PCR for BBa_B0030-BBa_C0061-BBa_B0030-BBa_C0078-BBa_B1006-BBa_R0079 plate: 6 colonies.
 |
- Gel results: all colonies were true positives! we decided to keep 1st colony to grow a 9 ml overnight culture.
- Plasmid digestion for:
| BBa_J23100-BBa_B0030-BBa_C0012-BBa_B1006 (E-S) | BBa_R0010 (E-X) |
- Gel run/cut/gel extraction.
- Ligation: BBa_J23100-BBa_B0030-BBa_C0012-BBa_B1006-BBa_R0010
- We incubated ligation at 16°C overnight.
- We infected 9 ml LB + Amp with 30 µl of these glycerol stocks:
| BBa_B0030-BBa_E0040-BBa_B1006 | BBa_B0030-BBa_C0061-BBa_B1006-BBa_R0062 |
| BBa_B0030-BBa_C0051-BBa_B0030-BBa_C0079-BBa_B1006-BBa_R0051-BBa_B0030-BBa_C0062-BBa_B1006-BBa_R0062 (1) | BBa_B0030-BBa_C0051-BBa_B0030-BBa_C0079-BBa_B1006-BBa_R0051-BBa_B0030-BBa_C0062-BBa_B1006 (1) |
| BBa_B0030-BBa_E1010-BBa_B1006 | BBa_B0030-BBa_C0078-BBa_B1006-BBa_R0079 |
| BBa_B0030-BBa_C0061-BBa_B0030-BBa_C0079-BBa_B1006-BBa_R0079 |
07/29/08
- Glycerol stocks/miniprep for the 7 overnight cultures. Unfortunately we had low yield for some of these parts...We are going to cut all these parts anyway.
- LB + Amp preparation.
- We transformed BBa_J23100-BBa_B0030-BBa_C0012-BBa_B1006-BBa_R0010 ligation. We plated transformed bacteria and incubated plate at 37°C overnight.
- We received sequencing results for:
- BBa_J23100-BBa_B0030-BBa_C0040-BBa_B1006-BBa_R0040
- BBa_B0030-BBa_C0078-BBa_B1006-BBa_R0079: sequence was correct!
07/30/08
- Plasmid digestion for:
| BBa_B0030-BBa_E0040-BBa_B1006 (X-P) | BBa_B0030-BBa_C0061-BBa_B1006-BBa_R0062 (S-P) |
| BBa_B0030-BBa_C0051-BBa_B0030-BBa_C0079-BBa_B1006-BBa_R0051-BBa_B0030-BBa_C0062-BBa_B1006-BBa_R0062 (1) (S-P) | BBa_B0030-BBa_C0061-BBa_B0030-BBa_C0079-BBa_B1006-BBa_R0079 (1) (S-P) |
| BBa_B0030-BBa_E1010-BBa_B1006 (X-P) | BBa_B0030-BBa_C0078-BBa_B1006-BBa_R0079 (S-P) |
- Gel run/cut/gel extraction. DNA quantification with NanoDrop showed that all cut parts are enough for the four ligations!
- Ligations:
- BBa_B0030-BBa_C0051-BBa_B0030-BBa_C0079-BBa_B1006-BBa_R0051-BBa_B0030-BBa_C0062-BBa_B1006-BBa_R0062-BBa_B0030-BBa_E1010-BBa_B1006
- BBa_B0030-BBa_C0061-BBa_B1006-BBa_R0062-BBa_B0030-BBa_E0040-BBa_B1006
- BBa_B0030-BBa_C0078-BBa_B1006-BBa_R0079-BBa_B0030-BBa_E0040-BBa_B1006
- BBa_B0030-BBa_C0061-BBa_B0030-BBa_C0079-BBa_B1006-BBa_R0079-BBa_B0030-BBa_E0040-BBa_B1006
- We incubated ligations at 16°C overnight.
