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

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