Team:TU Delft/23 July 2010 content
From 2010.igem.org
Alkane Degradation
With a number of yesterday's colonies a digestion was done to check if they contain the desired insert. The table below describes the digestion procedure, as well as the gel lane description.
# | Contents | Enzyme 1 | Enzyme 2 | Buffer | BSA | Expected size(s) (bp) |
1 | Smartladder (5μl) | n/a | ||||
2 | 007T (2) | 0.5 μl EcoRI | 0.5 μl PstI | 3 (Biolabs) | no | 1341, 2455 |
3 | 008T (4) | 0.5 μl EcoRI | 0.5 μl PstI | 3 (Biolabs) | no | 276, 2455 |
4 | 009T (6) | 0.5 μl EcoRI | 0.5 μl PstI | 3 (Biolabs) | no | 285, 2455 |
5 | 009T (7) | 0.5 μl EcoRI | 0.5 μl PstI | 3 (Biolabs) | no | 285, 2455 |
6 | 010T (9) | 0.5 μl EcoRI | 0.5 μl PstI | 3 (Biolabs) | no | 1382, 2455 |
7 | 017T (14) | 0.5 μl EcoRI | 0.5 μl PstI | 3 (Biolabs) | no | 855, 2455 |
8 | Empty | |||||
9 | 007T (2) | 1 μl PstI | 1 μl PvuII | H (Boehringer) | no | 2529,983,251 |
10 | 008T (4) | 1 μl AatII | 0.5 μl KpnI | A (Boehringer) | 1 μl | 271, 2427 |
11 | 009T (6) | 0.5 μl KpnI | 1 μl NruI | B (Boehringer) | 1 μl | 598, 2109 |
12 | 009T (7) | 0.5 μl KpnI | 1 μl NruI | B (Boehringer) | 1 μl | 598, 2109 |
13 | 010T (9) | 1 μl EarI | 0.5 μl PstI | 1 (Biolabs) | no | 1075, 2729 |
14 | 017T (14) | 0.5 μl AdhI | 0.5 μl SpeI | 4 (Biolabs) | no | 428, 2849 |
15 | EZ load (ladder) | n/a |
It seems (from the gel) that BioBrick 007T is the correct one, and the others don't contain the desired insert.