Team:Newcastle/8 July 2010

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Contents

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
  • 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.
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