0.5 ul destination vector
3 ul entry vector
2 ul ligation buffer
4 ul water
0.5 ul ligase
Incubate at room temp. for 1 hr or 4° over night
Thaw competent cells on ice (Be very gentle with these cells)
Add 3 ul of the ligation solution to the tube with the comp. cells, wiggle the drop down the side of the tube
Place on ice for 30 min
“Heat shock” the cells by placing them in 42° water bath for 20 sec.
Put on ice for 2 min
Add 250 ul pre-warmed S.O.C. solution
Shake at 37° for 1 hr.