Team:Warsaw/Calendar-Main/30 July 2009

Cloning of the mgtc promoter into the pSB1A3 plasmid Kamil

Tasks:  Colonies transfer Plasmid isolation and digest 

Methods:  Selected white colonies were transfered to a new dish, liquid cultures were established along the way. The liquid cultures were incubated for 6h at 37&deg;C with aeration. The plasmids were isolated according to the protocol (see at the end of the page). The plasmids were digested with EcoRI and PstI enzymes. The results were visualised with electrophoresis on 1% agarose gel.</li> NOTE: Steps 3 to 5 were not carried out by me and I cannot take credit for them. </ul>

Results:  Gel Electrophoresis:</li> </ul> <img src=http://2009.igem.org/wiki/images/7/79/2009.07.30.jpg /> From left:  GeneRuler DNA Ladder Mix #SM0333 (Fermentas)</li> Colonies from 1 to 10</li> GeneRuler DNA Ladder Mix #SM0333 (Fermentas)</li> </ul> NOTE: The upper band represents the plasmid backbone, the lower band is a bit odd. The mgtc promoter is roughly 500 bps in length while the only band except the backbone is somewhere around 1500 bps. Ligation picked up something completely wrong.

Conclusions:  Back to the drawing board, again... </li> </ul>

Construction of <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K177012 ">K177012</a>  operon1_part2 Ania Tasks:  Repeating the cloning process again and labeling my samples before going on holidays. Honestly I do not remember now what we did. </li>  Results: Not very promising so far </li></ul>