Team:Warsaw/Calendar-Main/12 May 2009

Lab meeting. PCR mgtc, pho Kama

Tasks:  Amplification of pho, mgtc 

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 (Salmonella) 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: pho 2min30s 95&deg;C (30s 95&deg;C, 35s 56&deg;C, 3min30s 72&deg;C)x3 (30s 95&deg;C, 35s 61&deg;C, 3min30s 72&deg;C)x28 10min 72&deg;C ~ 7&deg;C mgtc 1min30s 95&deg;C (30s 95&deg;C, 35s 48&deg;C, 1min 72&deg;C)x3 (30s 95&deg;C, 35s 58&deg;C, 1min 72&deg;C)x28 10min 72&deg;C ~ 7&deg;C  

Electrophoretic separation on 1% agarose gel Results:</P> <img src="http://2009.igem.org/wiki/images/2/27/2009.05.12_-_PCR_pho_i_mgtc_opisany.jpg"/>

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

Notes:  mgtc was succesfully amplified (termocykler z gradientem w pokoju 145)