July, 4th
Digestions of previously purified plasmids were performed for ligations:
| Plasmid |
Kind |
Purified Dna (μl) |
H2O (μl) |
Enzyme 1 (μl) |
Enzyme 2 (μl) |
Buffer H (μl) |
Final Volume(μl) |
| <partinfo>BBa_C0060</partinfo> |
Insert |
16.5 |
4 |
1 EcoRI |
1 SpeI |
2.5 |
25 |
| <partinfo>BBa_C0061</partinfo> |
Insert |
13.2 |
7.3 |
1 XbaI |
1 PstI |
2.5 |
25 |
| <partinfo>BBa_K081022</partinfo> |
Insert |
15.7 |
4.8 |
1 EcoRI |
1 PstI |
2.5 |
25 |
| <partinfo>BBa_B0030</partinfo> |
Vector |
13.2 |
7.3 |
1 SpeI |
1 PstI |
2.5 |
25 |
| <partinfo>BBa_B0031</partinfo> |
Vector |
12.4 |
8.1 |
1 SpeI |
1 PstI |
2.5 |
25 |
| <partinfo>BBa_B0032</partinfo> |
Vector |
9.5 |
11 |
1 SpeI |
1 PstI |
2.5 |
25 |
| <partinfo>BBa_B0015</partinfo> |
Vector |
7.9 |
12.6 |
1 EcoRI |
1 XbaI |
2.5 |
25 |
| <partinfo>pSB4C5</partinfo> |
Vector |
3 |
17.5 |
1 EcoRI |
1 PstI |
2.5 |
25 |
| <partinfo>BBa_I13501</partinfo> |
Insert |
7.2 |
13.3 |
1 XbaI |
1 PstI |
2.5 |
25 |
Reaction samples 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.
As shown in figure all clones were positive, so we cut and purified the bands of interest.
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