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9 July 2010

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

• Competent E. coli (DH5alpha strain)
• Ligation of lacI into pSB1AT3

Protocol

• Transformation of E. coli DH5α pSB1AT3 containing lacI insert.

Inference

• Ecoli DH5alpha is used as a bioreactor to replicate our plasmids to create a large stock ready for extraction from cells.