Team:Warsaw/Calendar-Main/23 July 2009

Cloning of mitochondrial targeting signal Sebastian Task 1:  PCR with gradient temperature of anniling  Methods:  Reaction mixture: 1ul pAcGFP1-Mito (5ng) 1ul dNTPs (2mM) 1ul L-primer (5pmol/ul) 1ul R-primer (5pmol/ul) 1ul Pfu buffer 4,5ul ddH20 0,5ul Pfu DNA polymerase  PCR program: 5min 95C 1min 95C 1min gradient 48-60C 1min 72C go to 2-nd step (30x) 5min 72C  

Cloning of the mgtc promoter into the pKSII+ plasmid Kamil

Tasks:  Transformant selection 

Methods:  The colonies that remained white after 48h were picked up and transferred to a new petri dish.</li> Liquid cultures were established along the way for plasmid purification.</li> Both the dish and the liquid cultures were incubated at 37&deg;C overnight.</li> </ul>

Assembly of endosomal detection operon Marcin Task 1:  Prepare the bacterial cultures for plasmid isolation</li> </ul> Preparation of bacterial cultures  Prepare LB medium with kanamycin</li> Add 3.5 ml of the medium to the probes</li> Add one bacterial colony to each probe</li> Breed the bacteria about 7 hours</li></ul>

Construction of <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K177012 ">K177012</a>  operon1_part2

Ania Tasks:

 Transformation of the chemocompetent cells with the ligation of <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_B0032"> BBa_B0032</a>- RBS.3 on the pSB1A2 ampicillin resistant plasmid and <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_C0012 BBa_C0012"> - lacI repressor</a> </li>