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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).

Screening for lacI insert:

1. Miniprep - the miniprep protocol was followed.
2. Restriction digests - the restriction digest protocol was followed.
   • Single digest with Pst1
   • double digests with Pst1 and Xba1
3. Gel electrophoresis

Set up liquid broth culture in 5 ml of LB - 1 colony from colonies 1-7