"
Team:Newcastle/9 July 2010
From 2010.igem.org
(Difference between revisions)
PhilipHall (Talk | contribs) (→Transformation) |
|||
| Line 1: | Line 1: | ||
{{Team:Newcastle/mainbanner}} | {{Team:Newcastle/mainbanner}} | ||
| - | + | '''9 July 2010''' | |
| - | === | + | =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). | * 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) | * Competent ''E. coli'' (DH5alpha strain) | ||
* Ligation of ''lacI'' into pSB1AT3 | * Ligation of ''lacI'' into pSB1AT3 | ||
| - | + | ==Protocol== | |
* [[TeamNewcastleTransformation of E. coli|Transformation of ''E. coli'']] DH5alpha pSB1AT3 containing ''lacI'' insert. | * [[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. | *''Ecoli'' DH5alpha is used as a bioreactor to replicate our plasmids to create a large stock ready for extraction from cells. | ||
{{Team:Newcastle/footer}} | {{Team:Newcastle/footer}} | ||
Revision as of 13:37, 11 August 2010
| |||||||||||||
| |||||||||||||
9 July 2010
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.
   
|
    
|
