Team:Warsaw/Calendar-Main/3 July 2009
From 2009.igem.org
Cloning of p53 coding sequence
Seth
Tasks:
- Restriction digest of p53 coding sequence obtained from 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°C
15 min 80°C
~4°C
Tasks:
- ligation of p53 coding sequence with pKS plasmid digested by XbaI and SmaI
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
Reaction occured about 18 h in 16°C
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