Team:Warsaw/Calendar-Main/3 July 2009

Cloning of p53 coding sequence Marcin Task 1:  Restriction digest of p53 coding sequence obtained from previously performed PCR reaction  Methods:   Reaction mixture composition: 15 ul PCR product (DNA concentration about 14.5 ng/ul) 5 ul Tango Buffer (Fermentas) 1 ul XbaI (Fermentas) 29 ul MQ water    digest program: digest 3h 37&deg;C 15 min 80&deg;C ~4&deg;C  

Task 2:  ligation of p53 coding sequence with pKS plasmid digested by XbaI and SmaI</li> </ul> Methods:   Ligation mixture composition: 15 ul digested p53 2 ul Tango Buffer (Fermentas) 1 digested pKS 0.5 ul ligase T4 2 ul 10 uM ATP </li> </ul>   Reaction was carried out about 18 h in 16&deg;C </li>