"
Team:Newcastle/8 July 2010
From 2010.igem.org
(Difference between revisions)
PhilipHall (Talk | contribs) (→Protocol) |
|||
| Line 1: | Line 1: | ||
{{Team:Newcastle/mainbanner}} | {{Team:Newcastle/mainbanner}} | ||
| - | |||
| - | === | + | '''8 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== | |
* Digested ''lacI'' | * Digested ''lacI'' | ||
* Excised gel containing pSB1AT3 | * Excised gel containing pSB1AT3 | ||
| - | + | ==Protocol== | |
* ''lacI'' digest is run on an [[Team:Newcastle/Gel_electrophoresis|agarose gel]] and excised | * ''lacI'' digest is run on an [[Team:Newcastle/Gel_electrophoresis|agarose gel]] and excised | ||
* Excised pSB1AT3 and ''lacI'' is then gel extracted | * Excised pSB1AT3 and ''lacI'' is then gel extracted | ||
* pSB1AT3 and ''lacI'' are [[Team:Newcastle/Ligation|ligated]] | * pSB1AT3 and ''lacI'' are [[Team:Newcastle/Ligation|ligated]] | ||
| - | + | ==Inference== | |
* ''lacI'' digest is purified by running on the agarose gel, removing unwanted fragments with homologous sticky ends. It is then ligated into vector pSB1AT3 ready to be transformed into ''E. Coli'' DH5α. | * ''lacI'' digest is purified by running on the agarose gel, removing unwanted fragments with homologous sticky ends. It is then ligated into vector pSB1AT3 ready to be transformed into ''E. Coli'' DH5α. | ||
| + | |||
{{Team:Newcastle/footer}} | {{Team:Newcastle/footer}} | ||
Revision as of 13:36, 11 August 2010
| |||||||||||||
| |||||||||||||
8 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
- Digested lacI
- Excised gel containing pSB1AT3
Protocol
- lacI digest is run on an agarose gel and excised
- Excised pSB1AT3 and lacI is then gel extracted
- pSB1AT3 and lacI are ligated
Inference
- lacI digest is purified by running on the agarose gel, removing unwanted fragments with homologous sticky ends. It is then ligated into vector pSB1AT3 ready to be transformed into E. Coli DH5α.
   
|
    
|
