Team:TU Delft/28 July 2010 content
From 2010.igem.org
Alkane degradation
The plates containing yesterday's ligations contained colonies, to check whether they really contain the desired BioBrick a colony PCR was done, and the used colonies were grown in liquid LB medium over night. The results from the PCR were analysed on a 1% agarose gel.
Lane Description part 1
# | Description | Expected lenght (bp) | Primers | Status |
1 | SmartLadder | n/a | n/a | n/a |
2 | J61100 + rubA3 | 473 | G00100 + G00101 | ✓ |
3 | J61100 + rubA3 | 473 | G00100 + G00101 | ✓ |
4 | J61100 + rubA3 | 473 | G00100 + G00101 | ✗ |
5 | J61100 + rubA3 | 473 | G00100 + G00101 | ✓ |
6 | J61100 + rubA3 | 473 | G00100 + G00101 | ✗ |
7 | J61100 + rubA4 | 482 | G00100 + G00101 | ✓ |
8 | J61100 + rubA4 | 482 | G00100 + G00101 | ✓ |
9 | J61100 + rubA4 | 482 | G00100 + G00101 | ✓ |
10 | J61100 + rubA4 | 482 | G00100 + G00101 | ✓ |
11 | J61100 + rubA4 | 482 | G00100 + G00101 | ✓ |
12 | J61100 + rubR | 1505 | G00100 + G00101 | ✓ |
13 | J61100 + rubR | 1505 | G00100 + G00101 | ✓ |
14 | J61100 + rubR | 1505 | G00100 + G00101 | ✓ |
15 | J61100 + rubR | 1505 | G00100 + G00101 | ✓ |
16 | J61100 + rubR | 1505 | G00100 + G00101 | ✓ |
17 | SmartLadder | n/a | n/a | n/a |
Lane Description part 2
# | Description | Expected lenght (bp) | Status | Remarks |
1 | SmartLadder | n/a | n/a | n/a |
2 | J61100 + ladA | 1625 | G00100 + G00101 | ✓ |
3 | J61100 + ladA | 1625 | G00100 + G00101 | ✓ |
4 | J61100 + ladA | 1625 | G00100 + G00101 | ✓ |
5 | J61100 + ladA | 1625 | G00100 + G00101 | ✓ |
6 | J61100 + ladA | 1625 | G00100 + G00101 | ✗ |
7 | J61101 + ADH | 1052 | G00100 + G00101 | ✓ |
8 | J61101 + ADH | 1052 | G00100 + G00101 | ✓ |
9 | J61101 + ADH | 1052 | G00100 + G00101 | ✗ |
10 | J61101 + ADH | 1052 | G00100 + G00101 | ✓ |
11 | J61101 + ADH | 1052 | G00100 + G00101 | ✗ |
12 | J61107 + ALDH | 1796 | G00100 + G00101 | ✗ |
13 | J61107 + ALDH | 1796 | G00100 + G00101 | ✓ |
14 | J61107 + ALDH | 1796 | G00100 + G00101 | ✗ |
15 | J61107 + ALDH | 1796 | G00100 + G00101 | ✗ |
16 | J61107 + ALDH | 1796 | G00100 + G00101 | ✗ |
17 | J61100 (uncut) | G00100 + G00101 | ✓ | |
18 | J61101 (uncut) | G00100 + G00101 | ✓ | |
19 | J61107 (uncut) | G00100 + G00101 | ✓ | |
20 | SmartLadder | n/a | n/a |
From the scPCR it can be concluded that from all the colonies PCRed (per BioBrick), at least one was positive! Next week the next (last) step in making the BioBricks for alkane degradation can be be done, so hopefully the week after that we can start characterizing.