Team:Macquarie Australia/Protocols/ligations


Ligation Procedure

A reaction mixture was prepared containing,

Element Volume
Upstream Digestion part 2 µL
Downstream Digestion part 2 µL
Destination Plasmid 1 µL
10X T4 DNA ligase buffer 2 µL
T4 DNA ligase 1 µL
H2O 12 µL

We incubated each ligation mix at 30°C for 30 minutes, followed by heat inactivation at 80°C for 20 minutes.

4 µL of the ligation product was transformed into 100 µL of competent E. coli, the transformants were then incubated for one hour.