First I cut my plasmid using restriction enzymes and ran this on agarose gel. I purified this, and all though the concentration is significantly lower I still have a certain amount of DNA. I perform mung bean nuclease to blunt the ends of the plasmid, and after following protocol and inactivating the enzymes by 0,01% SDS I run it on an agarose gel. The ladder migrates fine, but my DNA is still in the well. Why??
I would agree with Dr. Paul the SDS concetration might be pretty low . If you increase it and migration works properly perhaps is this. Might partially inactivation most likely woyld be the cause.
Best of Luck
Kat.
you may still have enzyme bound to the dna.Try dissociating with 0.5%sds and see if it helps
I would agree with Dr. Paul the SDS concetration might be pretty low . If you increase it and migration works properly perhaps is this. Might partially inactivation most likely woyld be the cause.
Best of Luck
Kat.
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