I isolated the my plasmids several times with isolation kits and to check , cut with restriction enzymes than run the agarose gel electrophoresis but ı got smear bands also background emission. What can be the reason(s) ?
I agree with Artur. The first gel looks fine, those smears aren't uncommon in enzyme digestions. The second gel is problematic because, in general, it looks pretty bad. Band intensities aren't great which makes it hard to interpret. Best to run on freshly prepared gels.
You could be over-digesting your plasmids, either by adding too much enzyme (or too little plasmid), or by letting the reactions incubate for too long. Make sure you're using the recommended concentrations of enzymes and the correct buffers for all.
Hi Ibrahim! I agree with Nathan and Artur. I can only say that any smear like that means that you have degraded material there. Using restrictions enzymes may produce a little bit of degradation, but If you put a lot of sample in the well, it may be more probable that the degraded DNA may be seen in a gel. I will try to run agin the sample with less amount. And about the background, it may be that one of your reagents is contaminated, Try to use a fresh batch of them.
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