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Team:Newcastle/16 July 2010
From 2010.igem.org
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*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 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 this 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. | + | *The aim of this 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 == | == Protocol == | ||
Revision as of 13:21, 6 August 2010
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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).
- The aim of this 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 gel electrophoresis protocol was followed.
Inference
- Set up liquid broth culture in LB
   
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