Team:Warsaw/Calendar-Main/17 September 2009

Cloning of the mgtc promoter into the pSB1A3 plasmid Kamil

Tasks:  Ligation to the plasmid Bacteria transformation 

Methods:  The ligation mix was prepared as follows: 20&mu;l purified plasmid backbone 20&mu;l mgtC promoter 5&mu;l LIgase Buffer (Fermentas) 1&mu;l T4 Ligase (Fermentas) 4&mu;l H2O  The ligation was carried out in 18&deg;C for 4h. A fresh batch of chemocompetent bacteria was transformed according to the protocol. 

Cloning of the cro-box into the pSB1A3 plasmid Kamil

Tasks:  Ligation to the plasmid</li> Bacteria transformation</li> </ul>

Methods:  The ligation mix was prepared as follows: 20&mu;l purified plasmid backbone 10&mu;l Cro-Box 4&mu;l LIgase Buffer (Fermentas) 5&mu;l 30% PEG 1&mu;l T4 Ligase (Fermentas)</li> The ligation was carried out in 18&deg;C for 4h.</li> A fresh batch of chemocompetent bacteria was transformed according to the protocol.</li> </ul>

Assembly of fusion proteins Marcin Task 1: Preparation of bacterial cultures to isolate mitochondrial peptide on pSB1A3 plasmid

One important remark:

Data from sequencing reactions clearly show that previously isolated plasmid which should contain: mitochondrial peptide CDS (BBa_K177028 - blunt ligation with pKSII) and BBa_K177037 are other commercial plasmids. Something is wrong - some plasmids samples have been contaminated.