Team:TU Delft/23 July 2010 content

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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 10μl end-volume)

# 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) 1341, 2455
3 008T (4) 0.5 μl EcoRI 0.5 μl PstI 3 (Biolabs) 276, 2455
4 009T (6) 0.5 μl EcoRI 0.5 μl PstI 3 (Biolabs) 285, 2455
5 009T (7) 0.5 μl EcoRI 0.5 μl PstI 3 (Biolabs) 285, 2455
6 010T (9) 0.5 μl EcoRI 0.5 μl PstI 3 (Biolabs) 1382, 2455
7 017T (14) 0.5 μl EcoRI 0.5 μl PstI 3 (Biolabs) 855, 2455
8 Empty
9 007T (2) 1 μl PstI 1 μl PvuII H (Boehringer) 2529,983,251
10 008T (4) 1 μl AatII 0.5 μl KpnI A (Boehringer) 271, 2427
11 009T (6) 0.5 μl KpnI 1 μl NruI B (Boehringer) 598, 2109
12 009T (7) 0.5 μl KpnI 1 μl NruI B (Boehringer) 598, 2109
13 010T (9) 1 μl EarI 0.5 μl PstI 1 (Biolabs) 1075, 2729
14 017T (14) 0.5 μl AdhI 0.5 μl SpeI 4 (Biolabs) 428, 2849
15 EZ load (ladder) n/a
Digestion check

It seems (from the gel) that BioBrick 007T is the correct one, and the others don't contain the desired insert.