Team:Warsaw/Calendar-Main/11 May 2009

PCR pho, mgtc, inv Kama

Tasks:  Amplification of pho, mgtc and 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) 1&mu;l template 0,5&mu;l pfu turbo polymerase (KNGiE) 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 inv 4min 95&deg;C (30s 95&deg;C, 1min 53&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/b/b8/2009.05.11_-_PCR_pho%2C_mgtc_i_inv_opisany.jpg"/>

 Gel (from left)</li> </ul> <ol> 1. GeneRuler DNA Ladder Mix #SM0333 (Fermentas)</li> 2. pho</li> 3. pho control -</li> 4. mgtc</li> 5. mgtc control -</li> 6. inv</li> 7. inv control -</li> </ol>

Notes:  No product. Check the template (Salmonella), polymerase (KNGiE) and dNTPs.</li> </ul>