"
Team:Newcastle/13 July 2010
From 2010.igem.org
(Difference between revisions)
PhilipHall (Talk | contribs) (→Materials and Protocol) |
|||
| Line 7: | Line 7: | ||
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 | + | ==Materials== |
| - | + | ||
| - | + | ||
| - | + | Seven [[Team:Newcastle/12_July_2010|overnight cultures]] | |
| - | + | Restriction enzyme ecoR1 | |
| - | ==== | + | ==Protocol== |
| - | *The [https://2010.igem.org/TeamNewcastleRestriction_digests| restriction | + | |
| + | *''E. coli'' DH5alpha was [https://2010.igem.org/TeamNewcastleTransformation_of_E._coli#Protocol|transformed]] again with our ligation mix. | ||
| + | *The seven overnight cultures underwent [[Team:Newcastle/Qiagen Minipreps|plasmid extraction]]. | ||
| + | *The seven plasmid extracts were then [https://2010.igem.org/TeamNewcastleRestriction_digests| restriction digested] with ecoR1 to linearise. | ||
| + | *The seven digests were then run on a gel. | ||
| + | |||
| + | ==Results== | ||
| + | This procedure was followed in order to determine whether our ''lacI'' had been ligated into the plasmid vector. The Gel shows that | ||
{{Team:Newcastle/footer}} | {{Team:Newcastle/footer}} | ||
Revision as of 09:15, 10 August 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
Seven overnight cultures Restriction enzyme ecoR1
Protocol
- E. coli DH5alpha was [1]] again with our ligation mix.
- The seven overnight cultures underwent plasmid extraction.
- The seven plasmid extracts were then restriction digested with ecoR1 to linearise.
- The seven digests were then run on a gel.
Results
This procedure was followed in order to determine whether our lacI had been ligated into the plasmid vector. The Gel shows that
   
|
    
|
