Restriction Enzyme Binding - (Aug/09/2007 )

Hi All,
Can restriction enzymes after digest stay bound to the plasmid they are cutting if not cleaned up properly? Any paper references on this?

I'm looking forward to your interesting responses. Thanks

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I haven't tested this but I don't think so... there are some RE that will bind to DNA and travel along the DNA strand looking for the appropriate target sequences, but even these will have an on/off rate... (like polymerase without the clamp the enzymes will fall off after processing for a little ways) So I would think that they do not stay bound.

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I think it is better to inactive enzymes by incubate it in 65 C for 30 min.Or with phenol .

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No papers for reference, but only our experience in the lab with such problems.

Heat inactivation for enzymes after digestion helps ligation as the enzymes donot interfere with the digested ends or with DNA while ligating it.

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