Team:Warsaw/Calendar-Main/3 July 2009
From 2009.igem.org
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°C
15 min 80°C
~4°C
Task 2:
- 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 was carried out about 18 h in 16°C
|
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
|
|