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Team:Newcastle/16 July 2010
From 2010.igem.org
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| - | *The single digests (lanes 1-7) and the double digests (lanes | + | *The single digests (lanes 1-7) and the double digests (lanes 9-15) are separated by the molecular marker (lane 8). |
Revision as of 10:23, 10 August 2010
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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).
- The aim of today's experiment is to screen for the lacI insert. This could be done using PCR but that would require specific primers and is not as reliable.
Protocol
- Miniprep - the miniprep protocol was followed.
- Restriction digests - the restriction digests protocol was followed to allow the length of the DNA to be checked using single and double digest.
- Single digest with Pst1
- double digests with Pst1 and Xba1
- Gel electrophoresis - the protocol for gel electrophoresis was followed.
- Set up liquid broth culture in LB. The protocol for growing an overnight culture was followed.
Results
- The single digests (lanes 1-7) and the double digests (lanes 9-15) are separated by the molecular marker (lane 8).
Figure 1: Gel electrophoresis of the single and double digests using Pst1 and Xba1.
Inference
   
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