We are using the plasmid standard curve method for real-time PCR (Taqman). We have noticed that recently our plasmids are degrading very quickly, to the point where the lowest copy number plasmids are undetectable, even if the plasmid is freshly made.
Does anyone have any idea why this might be happening?
The only thing we could come up with is that our old plasmids that don't degrade are expression plasmids where the insert has been ligated using restriction enzymes, whereas the newer, degrading plasmids are in TA cloning vectors.
Thanks for your help!
Degrading plasmid standards
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