Team:Newcastle/5 July 2010

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Contents 1 LacI BioBrick Construction 1.1 Aims 1.2 Materials 1.3 Protocol 1.4 Inference

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

• Refrigerated transformation plates.

Protocol

• Set up a liquid culture of colonies transformed with pMutin4 and pSB1AT3.

Inference

• Transformed Ecoli DH5α grown up overnight to create a large stock of plasmid ready for extraction.