Team:Warsaw/Calendar-Main/30 June 2009

PCR inv Kama

Tasks:  Amplification of inv 

Methods:   PCR mixture's composition: 2,5&mu;l pfu buffer (Fermentas) 2,5&mu;l MgSO4 (Fermentas) 1,5&mu;l primers 1&mu;l dNTPs (10 mM)(new) 1&mu;l template (Yersinia) 0,5&mu;l pfu turbo polymerase (KNGiE) 1&mu;l DMSO The solution was topped up with H2O to 25&mu;l.    PCR programs: inv 4min 95&deg;C (30s 95&deg;C, 1min 54&deg;C, 4min15s 72&deg;C)x31 10min 72&deg;C ~ 7&deg;C  

Electrophoretic separation on 1% agarose gel Results:</P> <img src="http://2009.igem.org/wiki/images/8/8f/2009_06_30_inv_opisany.jpg"/>

 Gel (from left)</li> </ul> <ol>  GeneRuler DNA Ladder Mix #SM0333 (Fermentas)</li>  inv</li>  inv control -</li> </ol>

Notes:  No product</li> </ul>

Cloning of mitochondrial targeting signal Sebastian

Tasks:  Transformation of E. coli DH5&alpha;</li>

Methods:

transformation of chemocompetent DH5&alpha; E.coli cells with 100ng of pAcGFP1-Mito Plating on LB medium supplemented with Kanamycin