Team:TU Delft/28 July 2010 content
From 2010.igem.org
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==Alkane degradation== | ==Alkane degradation== | ||
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[[Image:TUDelft_20100728_scPCR1.png|400px|thumb|left|1% agarose of colony PCR part 1. Gel runned at 100V for 1 hour. Of all samples 10 μL + 2 μL loadingbuffer was loaded. 5 μL was loaded of marker]] | [[Image:TUDelft_20100728_scPCR1.png|400px|thumb|left|1% agarose of colony PCR part 1. Gel runned at 100V for 1 hour. Of all samples 10 μL + 2 μL loadingbuffer was loaded. 5 μL was loaded of marker]] | ||
- | + | Lane Description part 1 | |
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[[Image:TUDelft_20100728_scPCR2b.png|400px|thumb|left|% agarose of colony PCR part 1. Gel runned at 100V for 1 hour. Of all samples 10 μL + 2 μL loadingbuffer was loaded. 5 μL was loaded of marker]] | [[Image:TUDelft_20100728_scPCR2b.png|400px|thumb|left|% agarose of colony PCR part 1. Gel runned at 100V for 1 hour. Of all samples 10 μL + 2 μL loadingbuffer was loaded. 5 μL was loaded of marker]] | ||
- | + | Lane Description part 2 | |
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Revision as of 16:07, 15 August 2010
Lab work
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.