Also, I tried setting up a reaction without enzyme. Again it got degraded. I have checked with different NEB buffers, MgCl2 the result is the same. I have autoclaved everything tip boxes, Eppendorfs, water to remove DNases contamination(if there) but, it's degrading. What could be the possible causes?
Sauvik Dasgupta
Check for the concentration as well as the quality of samples and make sure that it is not degraded.
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Andrew Cannon
I have a couple questions, for you. What do the lanes in your gel correspond to in terms of digestion? What is the source of your DNA and how large should it be once isolate? How is the DNA purified? Where is your ladder in the gel? And, how are is your gel run (ie percentage of agarose, voltage, time, buffer etc.)?
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