Team:TU Delft/28 July 2010 content

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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.

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

# Description Expected lenght (bp) Status Remarks
1 SmartLadder n/a n/a
2 J61100 + rubA3 473
3 J61100 + rubA3 473
4 J61100 + rubA3 473
5 J61100 + rubA3 473
6 J61100 + rubA3 473
7 J61100 + rubA4 482
8 J61100 + rubA4 482
9 J61100 + rubA4 482
10 J61100 + rubA4 482
11 J61100 + rubA4 482
12 J61100 + rubR 1505
13 J61100 + rubR 1505
14 J61100 + rubR 1505
15 J61100 + rubR 1505
16 J61100 + rubR 1505
17 SmartLadder n/a n/a
% 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

# Description Expected lenght (bp) Status Remarks
1 SmartLadder n/a n/a
2 J61100 + ladA 1625
3 J61100 + ladA 1625
4 J61100 + ladA 1625
5 J61100 + ladA 1625
6 J61100 + ladA 1625
7 J61101 + ADH 1052
8 J61101 + ADH 1052
9 J61101 + ADH 1052
10 J61101 + ADH 1052
11 J61101 + ADH 1052
12 J61107 + ALDH 1796
13 J61107 + ALDH 1796
14 J61107 + ALDH 1796
15 J61107 + ALDH 1796
16 J61107 + ALDH 1796
17 J61100 (uncut)
18 J61101 (uncut)
19 J61107 (uncut)
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.