Team:Newcastle/8 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
- Digested lacI
- Excised gel containing pSB1AT3
Protocol
- lacI digest is run on an agarose gel and excised
- Excised pSB1AT3 and lacI is then gel extracted and purified.
- pSB1AT3 and lacI are ligated
Inference
- lacI digest is purified by running on the agarose gel, removing unwanted fragments with homologous sticky ends. It is then ligated into vector pSB1AT3 ready to be transformed into E. Coli DH5α.