Metaphase Chromosome prep (NO2)

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.