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ligation - (Oct/04/2006 )

I am using Fermentas T4 ligase for ligation. I didn't inactivate the ligase before transformation. What will happen?


nothing. The different salt concentration or media that the transformation occurs in will pretty much inactivate the enzyme anyway. It may be more detrimental to have an active enzyme for subsequent digests or modifications.


Nothing will happe.
If you want to re-digest or sub-clon then you'll have a problem.


for electroporation, using too much raw ligation mix will cause arching.