Team:Warsaw/Calendar-Main/8 May 2009

PCR mgtc, pho Kama

Tasks:  Amplification of pho and 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) 0,5&mu;l pfu turbo polymerase (KNGiE) 1&mu;l template (Salmonella) The solution was topped up with H2O to 20&mu;l.

   PCR programs: pho 2min30s 95&deg;C (30s 95&deg;C, 35s 56&deg;C, 3min30s 72&deg;C)x3 (30s 58&deg;C, 35s 61&deg;C, 3min30 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 63&deg;C, 1min 72&deg;C)x28 10min 72&deg;C ~ 7&deg;C   Electrophoretic separation on 1% agarose gel 

Results: <img src="http://2009.igem.org/wiki/images/e/ef/2009.05.08_-_PCR_pho_i_mgtc_opisany.jpg"/>  Gel (from left)</li> </ul> <ol> GeneRuler DNA Ladder Mix #SM0333 (Fermentas)</li> pho</li> pho control -</li> mgtc</li> mgtc sample -</li>

</ol>

Notes:  denaturation temperature value was wrong for pho.</li> </ul>