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Team:TU Delft/23 July 2010 content
From 2010.igem.org
Surprise
We are very grateful for the help of our superior, Esengül in the lab. To thank her, we organized a surprise for her… In the afternoon a special delivery for Esengül arrived. Inside was a self-baked cake from a secret admirer. What a surprise this was! Half a day later, she found out that we were behind it and we all enjoyed eating the cake.
Lab work
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. (Digestion was done in 10 μL end-volume)
| # | Sample | Enzyme 1 | Enzyme 2 | Buffer | BSA | |
| 1 | 007T (2) | 0.5 μL EcoRI | 0.5 μL PstI | 3 (Biolabs) | ✗ | |
| 2 | 008T (4) | 0.5 μL EcoRI | 0.5 μL PstI | 3 (Biolabs) | ✗ | |
| 3 | 009T (6) | 0.5 μL EcoRI | 0.5 μL PstI | 3 (Biolabs) | ✗ | |
| 4 | 009T (7) | 0.5 μL EcoRI | 0.5 μL PstI | 3 (Biolabs) | ✗ | |
| 5 | 010T (9) | 0.5 μL EcoRI | 0.5 μL PstI | 3 (Biolabs) | ✗ | |
| 6 | 017T (14) | 0.5 μL EcoRI | 0.5 μL PstI | 3 (Biolabs) | ✗ | |
| 7 | 007T (2) | 1 μL PstI | 1 μL PvuII | H (Boehringer) | ✗ | |
| 8 | 008T (4) | 1 μL AatII | 0.5 μL KpnI | A (Boehringer) | ✓ | |
| 9 | 009T (6) | 0.5 μL KpnI | 1 μL NruI | B (Boehringer) | ✓ | |
| 10 | 009T (7) | 0.5 μL KpnI | 1 μL NruI | B (Boehringer) | ✓ | 598, 2109 |
| 11 | 010T (9) | 1 μL EarI | 0.5 μL PstI | 1 (Biolabs) | ✗ | |
| 12 | 017T (14) | 0.5 μL AdhI | 0.5 μL SpeI | 4 (Biolabs) | ✗ |
Lane description:
| # | Description | Expected Length (bp) | Status | Remarks | |
| M1 | SmartLadder | n/a | n/a | n/a | |
| 1 | 007T (2) + EcoRI + PstI | 1341, 2455 | |||
| 2 | 008T (4) + EcoRI + PstI | 276, 2455 | |||
| 3 | 009T (6) + EcoRI + PstI | 285, 2455 | |||
| 4 | 009T (7) + EcoRI + PstI | 285, 2455 | |||
| 5 | 010T (9) + EcoRI + PstI | 1382, 2455 | |||
| 6 | 017T (14) + EcoRI + PstI | 855, 2455 | |||
| 7 | Empty | ||||
| 8 | 007T (2) + PstI + PvuII | 2529,983,251 | |||
| 9 | 008T (4) + AatII + KpnI | 271, 2427 | |||
| 10 | 009T (6) + KpnI + NruI | 598, 2109 | |||
| 11 | 009T (7) + KpnI + NruI | 598, 2109 | |||
| 12 | 010T (9) + EarI + PstI | 1075, 2729 | |||
| 13 | 017T (14) + AdhI + SpeI | 428, 2849 | |||
| M2 | EZ load (ladder) |
It seems (from the gel) that BioBrick 007T is the correct one, and the others don't contain the desired insert.
