I was wondering if the nanodrop concentration readings changes as a result of RNA degradation in a certain sample which has been measured at two different time points, one straight after extraction and the other one a few months after? will the RNA concentration readings (ng/ul) increase, decrease or remains the same?
Yes - single ribonucleotides and deoxyribonucleotides give a stronger A260nm absorbance when no longer polymerized. The absorbance maxima for A, U, T, C and G are not all exactly 260nm... but nonetheless, when not polymerized, they all absorb more strongly at A260nm... See the IDT website and other basic biochemistry sources.
Hello Sharareh,
Nanodrop is a good tool for quantification and quality control of RNA via its maximum absorption at 260nm. But it's not a good tool to check integrity of RNA because degraded RNA absorb at the same level as intact RNA. So you shouldn't see difference if you measure RNA samples at different time point.
Best way to check intregrity after time is to perform RNA electrophoresis on agarose gel for example. You can find some of protocols on the web.
Hope it could help you.
Yes - single ribonucleotides and deoxyribonucleotides give a stronger A260nm absorbance when no longer polymerized. The absorbance maxima for A, U, T, C and G are not all exactly 260nm... but nonetheless, when not polymerized, they all absorb more strongly at A260nm... See the IDT website and other basic biochemistry sources.
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