Team:Newcastle/Ligation

For standard ligation 100-200ng of vector DNA is required - can be checked using nanodrop

A ligation calcultion can be used however a 1:3 and a 1:5 ratio tends to be used instead.

Set up ligation

Reagents	1:3(μl)	1:5(μl)	Vector(μl)
V	1	1	1
I	3	5
10*BUFFER	1	1	1
Ligase T4	1	1	1
H2O	4	2	7
Total Volume	10.0	10.0	10.0

For sticky end ligations incubate on the bench of 3-4 hrs or overnight