Team:Newcastle/13 July 2010
From 2010.igem.org
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Contents |
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 and Protocol
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.
Restriction digests
- The restriction digest protocol was followed for this step.