5 min rapid ligation protocol using 2X Quick Ligation Reaction Buffer
2X Quick Ligation Reaction Buffer:
132 mM Tris-HCl, pH 7.6 @25°C
20 mM MgCl2
2 mM DTT
2mM ATP
15% PEG (6000, 3350, or 8000).
Ligation:
10 ul DNA mix (insert + vector)
10 ul 2X ligase buffer
0.7 ul Ligase from NEB
Mix and ligate at RT for 5 min.
Transform bacteria
Ref: http://www.neb.com/nebecomm/products/productm2200.asp
