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).
To see whether the LacI insert is in the pSB1AT3 plasmid (same as yesterday).
Materials and Protocol
Performed a gel electrophoresis with remaining PCR products + Controls with same restriction digests.
Transformation
E. coli DH5alpha was transformed again with our plasmid. The transformation protocol was followed.
Miniprep
For this step the Qiagen Minipreps protocol was followed. The 7 colonies that grew on the plates from yesterday were cultured ovenight on a shaking shelf at 37°C in universal tubes.
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