Team:Newcastle/13 July 2010
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LacI BioBrick Construction
Aims
To use PCR to extract lacI (promoter, ribosome-binding site (RBS) & coding sequence (CDS)) from plasmid pMutin4 and ligate into vector pSB1AT3 in front of red fluorescent protein (RFP).
Materials
Seven overnight cultures Restriction enzyme ecoR1
Protocol
- E. coli DH5alpha was [[1]] again with our ligation mix.
- The seven overnight cultures underwent plasmid extraction.
- The seven plasmid extracts were then restriction digested with ecoR1 to linearise.
- The seven digests were then run on a gel.
Results
This procedure was followed in order to determine whether our lacI had been ligated into the plasmid vector. The Gel shows that