Trouble with AgeI digestion?

Hi all, I have been working on a cloning project and I think the reason it is failing is due to incomplete digestion. I am using EcoRI and AgeI sequentially, and am fairly certain that my EcoRI digestion is working, but I think AgeI is not cutting comletely. Currently, AgeI is the first digestion of the two that I run, I add ~5ug of DNA and 1.5uL of enzyme (7.5 units) and incubate for 2 hours at 37 degrees. Anyone had trouble with AgeI before? or have any tips for optimizing this digestion? Thanks!!

Posted Aug 14, 2017, 7:31 PM UTC

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