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Team:Newcastle/15 July 2010
From 2010.igem.org
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==Aims== | ==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). | *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'''. The experiments that were carried out yesterday are being repeated today as there were extra, unexpected bandings. | + | *The aim of today's experiment is to '''screen for the lacI insert'''. The experiments that were carried out [[Team:Newcastle/16_July_2010#Protocol|yesterday]] are being repeated today as there were extra, unexpected bandings. |
==Protocol== | ==Protocol== | ||
Revision as of 14:32, 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 today's experiment is to screen for the lacI insert. The experiments that were carried out yesterday are being repeated today as there were extra, unexpected bandings.
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.
Set up liquid broth culture in 5 ml of LB - 1 colony from colonies 1-7
   
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