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temp for ligation - (Mar/27/2006 )

i perform ligation with the insert and vector having blunt end in one side and salI cut in another side

i usually incubate the ligation sample in 16c . but is the temp is ok for blunt end also??

-T. reesei-

I did some blunt end ligations and I found that they worked best @ 4° C with a lot of PEG in the reactions, less ATP compared to sticky ends and more U of ligase.


I agree with genfranz. I actually never did a blunt end ligation, but i always use to ligate at 4°C .
Many buffers for blunt end ligation contain peg, so i would recommend it.
I also tried peg in sticky end ligation reactions, but i saw no significant difference.