Team:Newcastle/9 July 2010
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- | == | + | ==LacI BioBrick Construction== |
- | + | ===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). |
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===Materials=== | ===Materials=== | ||
- | + | * Competent ''E. coli'' (DH5alpha strain) | |
- | + | * Ligation of ''lacI'' into pSB1AT3 | |
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===Protocol=== | ===Protocol=== | ||
+ | * [[TeamNewcastleTransformation of E. coli|Transformation of ''E. coli'']] DH5alpha pSB1AT3 containing ''lacI'' insert. | ||
- | + | ===Inference=== | |
- | + | *''Ecoli'' DH5alpha is used as a bioreactor to replicate our plasmids to create a large stock ready for extraction from cells. | |
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{{Team:Newcastle/footer}} | {{Team:Newcastle/footer}} |
Revision as of 12:24, 4 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).
Materials
- Competent E. coli (DH5alpha strain)
- Ligation of lacI into pSB1AT3
Protocol
- Transformation of E. coli DH5alpha pSB1AT3 containing lacI insert.
Inference
- Ecoli DH5alpha is used as a bioreactor to replicate our plasmids to create a large stock ready for extraction from cells.