T4 ligase - is the inactivation step necessary? - (Mar/22/2005 )
I am using T4 ligase from invitrogen. the product manual said after incubation add 1ul 0.5MEDTA to inactivate the ligase. I did not do this before, do you do this inactivation step?
i never do this. After ligation i pellet the dna using ethanol/na acetate and electropore directly.
I use the ligation mix directly for transformation
yep, I just use the ligation reaction for transformation. It has worked for me everytime.
ethanol precipitate and transform.