Team:TU Delft/2 August 2010 content
From 2010.igem.org
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|SpeI | |SpeI | ||
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|‘E – J61100-rubR – S’ | |‘E – J61100-rubR – S’ | ||
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|‘E – B0015 – pSB1AK3 – X’ | |‘E – B0015 – pSB1AK3 – X’ | ||
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Revision as of 09:19, 3 August 2010
Alkane Sensing, Solvent Tolerance and Salt Tolerance
by Pieter
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.
Alkane degradation
For the next step in BioBrick formation a digestion was done: Digestion Some BioBricks are in production. Digestion reactions:
# | Sample | Enzyme 1 | Enzyme 2 | Enzyme 3 | Buffer | BSA | Needed fragment |
1 | 1 μg 007T | EcoRI | SpeI | BamH1 | 2 (Biolabs) | ✓ | ‘E – J61100-AlkB2 – S’ |
2 | 1 μg 008A | EcoRI | XbaI | 2 (Biolabs) | ✓ | ‘E – J61100-rubA3 – X’ | |
3 | 1 μg 010A | EcoRI | SpeI | AseI | 2 (Biolabs) | ✓ | ‘E – J61100-rubR – S’ |
4 | 2 μg B0015 | EcoRI | XbaI | 2 (Biolabs) | ✓ | ‘E – B0015 – pSB1AK3 – X’ | |
5 | 1 μg 017A | EcoRI | SpeI | AseI | 2 (Biolabs) | ✓ | ‘E – J61100-ladA – S’ |
6 | 1 μg 018A | XbaI | PstI | AseI | 2 (Biolabs) | ✓ | ‘X – J61101-ADH – P’ |
7 | 1 μg 019A | EcoRI | SpeI | AseI | 2 (Biolabs) | ✓ | ‘E – J61107-ALDH – S’ |
8 | 1 μg pSB1K3 | EcoRI | PstI | 2 (Biolabs) | ✓ | ‘E – pSB1K3 – P’ |
These