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Old 12-02-2016, 06:42 AM   #1
dlv04c
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Default Bioanalyzer RNA Nano with total RNA - High MW peak above 28S?

I'm getting a significant high MW peak on a nano RNA chip with total RNA. Any thoughts on what this might be caused by? It is present in all the total RNA samples I loaded on this chip.


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Old 12-06-2016, 11:39 PM   #2
Amar
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Possibly genomic DNA. Have you digested your DNA and if so with what?

Some good info here:
http://www2.jabsom.hawaii.edu/genomi...rpretation.pdf
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Old 12-07-2016, 10:40 AM   #3
dlv04c
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Default it was not denatured

Turns out the RNA was not denatured prior to loading on the chip.
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Old 10-24-2017, 07:38 AM   #4
marta.m_
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Did it help? You don't see this peak anymore?
I also have problem with this peak (though it is a bit smaller in my case). I am always denaturing my RNA prior to the run but maybe 2 min is not enough?

I think it is not DNA as second DNase treatment did not help and qPCR (+RT vs -RT, housekeeping gene) shows that the RNA is free of DNA (no amplification after 40 cycles). I also tried spiking in a small amount of gDNA and it never resulted in such electropherogram.


Then again I have seen this peak in many publications and examples of "good RNA".
http://www.nature.com/app_notes/nmet...eth908_F1.html
http://genomics.nchresearch.org/submitting_RNA.html
http://www.sigmaaldrich.com/technica...ssessment.html fig 7.9
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4535892/ fig 1.

What can that be?
I am very puzzled.
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