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Team:TU Delft/12 July 2010 content
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|<b>Composed of: </b> | |<b>Composed of: </b> | ||
|- | |- | ||
| - | |[http://partsregistry.org/Part: | + | |[http://partsregistry.org/Part:BBa_K398500 K398500] |
|J23100 + J61100 + I13401 | |J23100 + J61100 + I13401 | ||
|- | |- | ||
| - | |[http://partsregistry.org/Part: | + | |[http://partsregistry.org/Part:BBa_K398501 K398501] |
|J23100 + J61101 + I13401 | |J23100 + J61101 + I13401 | ||
|- | |- | ||
| - | |[http://partsregistry.org/Part: | + | |[http://partsregistry.org/Part:BBa_K398502 K398502] |
|J23100 + J61107 + I13401 | |J23100 + J61107 + I13401 | ||
|- | |- | ||
| - | |[http://partsregistry.org/Part: | + | |[http://partsregistry.org/Part:BBa_K398503 K398503] |
|J23100 + J61117 + I13401 | |J23100 + J61117 + I13401 | ||
|- | |- | ||
| - | |[http://partsregistry.org/Part: | + | |[http://partsregistry.org/Part:BBa_K398504 K398504] |
|J23100 + J61127 + I13401 | |J23100 + J61127 + I13401 | ||
|} | |} | ||
Revision as of 17:56, 15 July 2010
Lab Work
Characterization of Anderson RBS sequences
E.coli TOP 10 strains containing the following composite biobricks in pSB1A2 had been obtained:
| BioBrick # | Composed of: |
| [http://partsregistry.org/Part:BBa_K398500 K398500] | J23100 + J61100 + I13401 |
| [http://partsregistry.org/Part:BBa_K398501 K398501] | J23100 + J61101 + I13401 |
| [http://partsregistry.org/Part:BBa_K398502 K398502] | J23100 + J61107 + I13401 |
| [http://partsregistry.org/Part:BBa_K398503 K398503] | J23100 + J61117 + I13401 |
| [http://partsregistry.org/Part:BBa_K398504 K398504] | J23100 + J61127 + I13401 |
To prepare for the fluorescence assay measurements the strains were grown in M9 minimal medium containing 0.4% glucose and ampicillin.
High copy number RBS characterizations are rarely indicative of RiPS in system operation plasmids, thus the fluorescence assays will also be performed with the biobricks on pSB3C5. In order to obtain sufficient BioBrick DNA the strains were grown in 250 mL LB-Amp, awaiting QIAGEN Midi kit plasmid isolation and plasmid swapping.
Emulsifier The bricks for emulsifier production were assembled. The stock plasmids containing AlnA, OprG, R0011 and B0032 were digested and ligated into plasmid pSB1T3.
Digestion substrates:
| Label | Content | Concentration (ng/ul) |
| D1 | Plasmid [http://partsregistry.org/Part:pSB1T3 pSB1T3] | 137.2 |
| D2 | AlnA | 357.4 |
| D3 | OprG | 197.3 |
| D4 | B0015 | 360.5 |
| D5 | R0011 | 42.0 |
| D6 | B0032 | 82.0 |
Digested according to the digestion protocol (Link needed!).
The digestion products were ligated over night to produce the following biobricks:
| Label | BioBrick # | Composed of: |
| L1 | Control | Plasmid |
| L2 | [http://partsregistry.org/Part:BBa_K398202 K398202] | R0011 + B0032 |
| L3 | [http://partsregistry.org/Part:BBa_K398203 K398203] | OprG + B0015 |
| L4 | [http://partsregistry.org/Part:BBa_K398204 K398204] | AlnA + B0015 |
