Team:TU Delft/20 September 2010 content
From 2010.igem.org
Alkane degradation
For some reason, 009 and 010 just don't want to get into one plasmid. As a last resort we'll try every possible way to get the two together.
Digestions:
Lane # | BioBrick | Cut with | Extra | Buffer | BSA | Gel ok? |
0 | Smartladder (5μl) | |||||
D 1 | 009A | EcoRI, SpeI | AseI | Buffer 2 (NEB) | 1x BSA | OK |
D 2 | 009A | SpeI, PstI | Phosphatase | Buffer 2 (NEB) | 1x BSA | OK |
D 3 | 009A | SpeI, PstI | none | Buffer 2 (NEB) | 1x BSA | OK |
D 4 | 009A | EcoRI, XbaI | none | Buffer 2 (NEB) | 1x BSA | OK |
D 5 | 009A | XbaI, PstI | AseI | Buffer 2 (NEB) | 1x BSA | OK |
D 6 | 010A | EcoRI, SpeI | AseI | Buffer 2 (NEB) | 1x BSA | OK |
D 7 | 010A | SpeI, PstI | none | Buffer 2 (NEB) | 1x BSA | OK |
8 | 010A | EcoRI, XbaI | Phosphatase | Buffer 2 (NEB) | 1x BSA | OK |
D 9 | 010A | EcoRI, XbaI | none | Buffer 2 (NEB) | 1x BSA | OK |
D 10 | 010A | XbaI, PstI | AseI | Buffer 2 (NEB) | 1x BSA | OK |
Ligations:
'# | 'Piece 1 | Piece 2 |
028 L1 | D1 | D8 |
028 L2 | D1 | D9 |
028 L3 | D2 | D10 |
028 L4 | D3 | D10 |
028 L5 | D4 | D6 |
028 L6 | D5 | D7 |