In DNA gel electrophoresis my plasmid DNA is above DNA ladder which indicates nicked dna.
Plasmid was purified by plasmid miniprep kit.
My plasmid is not digesting. Enzymes are fine.
Agarose gel 1.5%
Kannan Balakrishnan
vector pET28a+ (concentration 150ng).enzymes Xba1 and Xho1. Gene of interest size 869bp. enzymes are digesting other vectors successfully.
other enzymes are also not digesting. such as nde1.
Nicked plasmid should run with the mobility approximately the same as single cut plasmid. In your case pET28 with that insert should be about 6kb.
I think the main problem is that a 1.5% gel is much too high percentage for this size range. Around 0.7% would be much better. You won't get good resolving of larger bands in a 1.5%. So I'm not really sure what you are looking at in these gels.
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