Team:Newcastle/13 July 2010
From 2010.igem.org
(Difference between revisions)
PhilipHall (Talk | contribs) (→Materials and Protocol) |
PhilipHall (Talk | contribs) (→Protocol) |
||
Line 14: | Line 14: | ||
==Protocol== | ==Protocol== | ||
- | *''E. coli'' DH5alpha was [https://2010.igem.org/TeamNewcastleTransformation_of_E._coli#Protocol|transformed]] again with our ligation mix. | + | *''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 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 plasmid extracts were then [https://2010.igem.org/TeamNewcastleRestriction_digests| restriction digested] with ecoR1 to linearise. |
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