Brief question:

I have performed a double digest with heat inactivated restriction enzymes with no star activity. I usually purify the DNA (PCR inserts and linearised vector) using a kit or gel electrophoresis but today I don't have time for it.

Is it okay to freeze (-20*C) the PCR inserts and linearised vector following heat inactivation of the enzymes? After a week, I will thaw, purify and perform ligation on these products.



From my experience I would say that it is no problem to freeze the digested DNA, when the enzyme is inactivated (even when not, for this time period I wouldn't expect much damage).

However, if possible, I would rather opt to freeze the PCR product and make the digest fresh, since the overhanging ends are kind of sensitive to hydrolytic degradation. I have also frozen linearized vectors without problems.


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