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Team:UNIPV-Pavia/Notebook/Week7
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*glycerol stocks. | *glycerol stocks. | ||
| + | |||
| + | <br><br> | ||
| + | '''07/3/08''' | ||
| + | <br> | ||
| + | *All the ligation plates showed carpets, while negative control plates showed a weak background noise. | ||
| + | |||
| + | *Single colonies plates for ligation plates. | ||
| + | |||
| + | *Glycerol stocks for: | ||
| + | {|cellpadding="20px" | ||
| + | |BBa_B0030 | ||
| + | |'''BBa_B0030'''-BBa_E0040 | ||
| + | |'''BBa_B0030'''-BBa_C0051 | ||
| + | |- | ||
| + | |BBa_B1006 | ||
| + | |'''BBa_B0030'''-BBa_E1010 | ||
| + | |} | ||
| + | |||
| + | *Miniprep for these parts. | ||
| + | |||
| + | *We performed digestion: | ||
| + | {|cellpadding="20px" | ||
| + | |BBa_B0030 (E-X) | ||
| + | |'''BBa_B0030'''-BBa_E0040 (E-S) | ||
| + | |'''BBa_B0030'''-BBa_C0051 (E-S) | ||
| + | |- | ||
| + | |BBa_B1006 (E-X) | ||
| + | |'''BBa_B0030'''-BBa_E1010 (E-S) | ||
| + | |} | ||
| + | |||
| + | *Gel run/cut for: | ||
| + | {|cellpadding="20px" | ||
| + | |'''BBa_B0030'''-BBa_E0040 (E-S) | ||
| + | |'''BBa_B0030'''-BBa_C0051 (E-S) | ||
| + | |'''BBa_B0030'''-BBa_E1010 (E-S) | ||
| + | |} | ||
| + | |||
| + | *Gel extraction. | ||
| + | |||
| + | *DNA precipitation with sodium acetate for: | ||
| + | {|cellpadding="20px" | ||
| + | |BBa_B0030 (E-X) | ||
| + | |BBa_B1006 (E-X) | ||
| + | |} | ||
| + | |||
| + | Ligation: | ||
| + | **BBa_B0030-BBa_E0040-'''BBa_B1006''' | ||
| + | **BBa_B0030-BBa_C0051-'''BBa_B0030''' | ||
| + | **BBa_B0030-BBa_E1010-'''BBa_B1006''' | ||
| + | |||
| + | *We incubated ligation reactions at 16°C overnight. | ||
| + | |||
| + | <br><br> | ||
| + | '''07/4/08''' | ||
| + | <br> | ||
| + | *We transformed the 3 ligations (2 µl) and plated transformed bacteria. | ||
| + | |||
| + | *Colony PCR for single colonies plates (3 colonies for each plate): | ||
| + | {|cellpadding="20px" | ||
| + | |BBa_J23100-'''BBa_B0030'''-BBa_C0012 | ||
| + | |BBa_J23100-'''BBa_B0030'''-BBa_C0040 | ||
| + | |BBa_J23100-'''BBa_B0030'''-BBa_I15010 | ||
| + | |- | ||
| + | |'''BBa_R0051'''-BBa_B0030-BBa_C0062 | ||
| + | |'''BBa_B0030'''-BBa_C0061-BBa_B1006 | ||
| + | |} | ||
Revision as of 22:17, 13 July 2008
 Home
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|
|---|---|---|---|---|
 Dry Lab
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|  Protocols
|  Activity Notebook
|
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 7: 06/30/08 - 06/5/08
06/30/08
- We received sequencing results for BBa_B0030-BBa_C0061 (4th colony and 7th colony): sequences were correct! We decided to keep the 4th colony.
- Colony PCR for BBa_J23100-BBa_E0240 and BBa_B0030-BBa_C0061: 10 colonies for every plate. Gel showed many working colonies: we chose first colonies for the two ligations.
 |
- We infected 9 ml LB + Amp with 30 µl of:
| BBa_C0012 | BBa_B1006 | BBa_R0051-BBa_B0030 | BBa_B0030-BBa_C0061 |
| BBa_C0062 | BBa_J23100-BBa_E0240 (1) | BBa_B0030-BBa_C0078 (1) | BBa_J23100-BBa_B0030 |
| BBa_C0040 | BBa_I15010 |
- Tomorrow we will be ready to perform 5 ligations!
07/1/08
- Glycerol stocks for:
| BBa_C0012 | BBa_B1006 | BBa_R0051-BBa_B0030 | BBa_B0030-BBa_C0061 |
| BBa_C0062 | BBa_J23100-BBa_E0240 (1) | BBa_B0030-BBa_C0078 (1) | BBa_J23100-BBa_B0030 |
| BBa_C0040 | BBa_I15010 |
- Miniprep for all these parts.
- We sent BBa_J23100-BBa_E0240 (1) and BBa_B0030-BBa_C0078 (1) to Primm for sequencing.
- We performed digestion for:
| BBa_C0012 (X-P) | BBa_B1006 (E-X) | BBa_R0051-BBa_B0030 (S-P) | BBa_B0030-BBa_C0061 (E-S) |
| BBa_C0062 (X-P) | BBa_C0040 (X-P) | BBa_I15010 (X-P) | BBa_J23100-BBa_B0030 (S-P) |
- Gel run/cut for:
| BBa_C0012 (X-P) | BBa_C0062 (X-P) | BBa_B0030-BBa_C0061 (E-S) |
| BBa_C0040 (X-P) | BBa_I15010 (X-P) |
- Gel extraction for these 5 parts.
- DNA precipitation with sodium acetate for:
| BBa_B1006 (E-X) | BBa_R0051-BBa_B0030 (S-P) | BBa_J23100-BBa_B0030 (S-P) |
- Ligation:
- BBa_J23100-BBa_B0030-BBa_C0012
- BBa_J23100-BBa_B0030-BBa_C0040
- BBa_J23100-BBa_B0030-BBa_I15010
- BBa_R0051-BBa_B0030-BBa_C0062
- BBa_B0030-BBa_C0061-BBa_B1006
- We incubated ligation reaction at 16°C overnight.
07/2/08
- We transformed ligations (5 µl) and plated transformed bacteria.
- We also transformed BBa_B1006(E-X), BBa_J23100-BBa_B0030(S-P) and BBa_R0051-BBa_B0030 (1 µl) and plated transformed bacteria to estimate background noise.
- We received sequencing results for:
- BBa_B0030-BBa_E0040
- BBa_B0030-BBa_C0051
- BBa_B0030-BBa_E1010
- All the sequences were correct!
- We infected 9 ml LB + Amp with 30 µl of:
| BBa_B0030 | BBa_B0030-BBa_E0040 | BBa_B0030-BBa_C0051 |
| BBa_B1006 | BBa_B0030-BBa_E1010 |
- glycerol stocks.
07/3/08
- All the ligation plates showed carpets, while negative control plates showed a weak background noise.
- Single colonies plates for ligation plates.
- Glycerol stocks for:
| BBa_B0030 | BBa_B0030-BBa_E0040 | BBa_B0030-BBa_C0051 |
| BBa_B1006 | BBa_B0030-BBa_E1010 |
- Miniprep for these parts.
- We performed digestion:
| BBa_B0030 (E-X) | BBa_B0030-BBa_E0040 (E-S) | BBa_B0030-BBa_C0051 (E-S) |
| BBa_B1006 (E-X) | BBa_B0030-BBa_E1010 (E-S) |
- Gel run/cut for:
| BBa_B0030-BBa_E0040 (E-S) | BBa_B0030-BBa_C0051 (E-S) | BBa_B0030-BBa_E1010 (E-S) |
- Gel extraction.
- DNA precipitation with sodium acetate for:
| BBa_B0030 (E-X) | BBa_B1006 (E-X) |
Ligation:
- BBa_B0030-BBa_E0040-BBa_B1006
- BBa_B0030-BBa_C0051-BBa_B0030
- BBa_B0030-BBa_E1010-BBa_B1006
- We incubated ligation reactions at 16°C overnight.
07/4/08
- We transformed the 3 ligations (2 µl) and plated transformed bacteria.
- Colony PCR for single colonies plates (3 colonies for each plate):
| BBa_J23100-BBa_B0030-BBa_C0012 | BBa_J23100-BBa_B0030-BBa_C0040 | BBa_J23100-BBa_B0030-BBa_I15010 |
| BBa_R0051-BBa_B0030-BBa_C0062 | BBa_B0030-BBa_C0061-BBa_B1006 |
