July, 25th
Red colonies were observed from E36 plates, suggesting that the transformation was successful. A colony was picked form the plate.
Digestions of previously purified plasmids were performed for ligations:
| Plasmid |
Kind |
DNA (μl) |
H2O (μl) |
Enzyme 1 (μl) |
Enzyme 2 (μl) |
Buffer H (μl) |
Final Volume (μl) |
| E24-2 |
Insert |
13 |
7.5 |
1 Xbal |
1 Pstl |
2.5 |
25 |
| E25-1 |
Insert |
13.5 |
7 |
1 Xbal |
1 Pstl |
2.5 |
25 |
| E26-2 |
Insert |
14.5 |
6 |
1 Xbal |
1 Pstl |
2.5 |
25 |
| E27-1 |
Insert |
12.5 |
8 |
1 Xbal |
1 PstI |
2.5 |
25 |
Digestions of previously purified plasmids (carrying either E17, or E18, or E19 or E20 part) were performed for ligations:
| Plasmid |
Kind |
DNA (μl) |
H2O (μl) |
Enzyme 1 (μl) |
Enzyme 2 (μl) |
Buffer H (μl) |
Final Volume (μl) |
| E17 |
Insert |
- |
- |
1 Xbal |
1 Pstl |
2.5 |
25 |
| E18 |
Insert |
- |
- |
1 Xbal |
1 Pstl |
2.5 |
25 |
| E19 |
Insert |
- |
- |
1 Xbal |
1 Pstl |
2.5 |
25 |
| E20 |
Insert |
- |
- |
1 Xbal |
1 PstI |
2.5 |
25 |
Reactions were incubated at 37°C for three hours while a small-size and a medium-size agarose gel were prepared according to protocols.
In the afternoon gel electrophoresis was performed.
Immagine della corsa su gel
After gel extraction, digested DNA was quantified:
| Part |
DNA (ng/μl) |
| E24 (E-P) |
- |
| E25 (E-P) |
- |
| E26 (E-P) |
- |
| E27 (E-P) |
- |
"
