"

From 2010.igem.org

Ligation

Ligations (pasting plasmid DNA) were performed at the appropriate temperature with the appropriate buffer in the appropriate concentration, according to the supplier.

Reaction for one sample:

DNA insert	x μL
DNA vector	x μL (for 1×)
T4 Ligation buffer (10×)	2.5 μL (for 1×)
T4 Ligase	x μL
H2O	x μL
	25,0 μL (final contration ~100 ng/μL)

Transform circa half of the ligation mix. Incubate at 16 °C o/n