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Team:UNIPV-Pavia/Notebook/Week12
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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: 08/4/08 - 08/8/08
08/4/08
- Plasmid digestion for:
| BBa_J23100-BBa_B0030-BBa_C0040-BBa_B1006 (E-S) | BBa_R0040 (E-X) |
- Gel run/cut/gel extraction.
- Ligation: BBa_J23100-BBa_B0030-BBa_C0040-BBa_B1006-BBa_R0040. We incubated ligation at 16°C overnight.
- We had 5 plates to screen with colony PCR:
- 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 (that we call "a")
- BBa_B0030-BBa_C0061-BBa_B1006-BBa_R0062-BBa_B0030-BBa_E0040-BBa_B1006 (that we call "b")
- BBa_B0030-BBa_C0078-BBa_B1006-BBa_R0079-BBa_B0030-BBa_E0040-BBa_B1006 (that we call "c")
- BBa_B0030-BBa_C0061-BBa_B0030-BBa_C0079-BBa_B1006-BBa_R0079-BBa_B0030-BBa_E0040-BBa_B1006 (that we call "d")
- BBa_J23100-BBa_B0030-BBa_C0040-BBa_B1006-BBa_R0040
- Last week BBa_J23100-BBa_B0030-BBa_C0040-BBa_B1006-BBa_R0040 colony PCR gave a bad result. For this reason, we decided to perform colony PCR only for:
- BBa_B0030-BBa_C0061-BBa_B1006-BBa_R0062-BBa_B0030-BBa_E0040-BBa_B1006 (7 colonies)
- BBa_J23100-BBa_B0030-BBa_C0012-BBa_B1006-BBa_R0010 (6 colonies)
 |
- Gel result:
- b (2nd colony, but it was not pure. We decided to prepare single colonies plate for b)
- BBa_J23100-BBa_B0030-BBa_C0012-BBa_B1006-BBa_R0010 (1st colony)
08/4/08
- Plasmid digestion for:
| BBa_J23100-BBa_B0030-BBa_C0040-BBa_B1006 (E-S) | BBa_R0040 (E-X) |
- Gel run/cut/gel extraction.
- We infected 9 ml of LB + Amp with 30 µl of BBa_C0062, 12, 22, 30, 27(2nd col), b(1st col)
- Single colonies plates for b, c, d.
08/4/08
- Glycerol stocks/miniprep for BBa_C0062, 12, 22, 30, 27(2), b(1).
- We sent BBa_C0062, 12, 22 and 30 to Primm for sequencing: all these parts contain BBa_C0062.
- We transformed/plated 28.
- Colony PCR for a(7 colonies), b(single colonies)(6 colonies), c(single colonies)(6 colonies), d(single colonies)(6 colonies).
- Gel results: PCR
