Blunt-Cohesive ligation - Should I change the conditions? (Aug/20/2010 )

Hi guys!

One quick question. When you perform a ligation with both ends (Blunt and cohesive), do you add PEG to the reaction and do you increase the amount of ligase?

Thanks!

Hi,
i performed ligation with cohesive ends without using PEG.the enzyme was T4 DNA ligase. but my friend who performed ligation with blunt ends use PEG to improve final efficiancy.

maral on Fri Aug 20 09:37:51 2010 said:

yeah sure, but I want to know when you have both ends how you do it. I have done them before without PEG and without add more ligase, I just want to know how you guys do it.

thanks