Cloning of p53 coding sequence

Marcin

Tasks:

• Transformation of chemocompetent E. coli strain DH5α

Comment:


When we found have appropriate bacteria I transformed it using the ligation reaction prepared in 04.07.09 to create pKS plasmid with p53 coding sequence

Methods:

• thaw bacteria on the ice - 10 minuts
• add 4 ul of ligation mixture to the bacteria
• incubation on the ice - 20 minutes
• heat shock - 1 minut, 42°C
• incubation on the ice - 2 minuts
• add 800 ul of SOB medium to the bacteria
• incubation in 37°C - 1 h
• Plating on selective LB medium supplemented with ampicillin, X-Gal and IPTG