Blunt Ligation Reaction

Azzy · #1 (**permalink**) 04-24-2008, 12:15 PM

What happens when the HindIII-EcoRI DNA is ligated by T4 DNA ligase? If the DNA digest fragments were blunt-ended how would you have changed the ligation reaction?

admin · #2 (**permalink**) 04-25-2008, 08:16 AM

Hell Azzy,
Blunt ends are generally the same but they usually are less efficient. Thats why most people make sure to keep the DNA ligation at 16C the most efficient temperature for the enzyme, and also to increase the time for ligation to overnight or even use high efficiency DNA ligases which are on sale for blunt end ligations.

Please let us know how you blunt ended the hindiii and ecori, and also how you prepared the vector as that is quite important too.

cheers!

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Blunt Ligation Reaction

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