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Old 03-28-2017, 11:48 AM   #9
eingabe_ausgabe
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Location: sacramento, ca

Join Date: Mar 2017
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Quote:
Originally Posted by Olaf Blue View Post
Have you tried depleting the rRNA using Ribo-Zero or RNAse H methods and seeing if or how many of the small RNAs remain or are eliminated using Bioanalyzer?
No and it may be too late for me to do that. I am actually trying to retain the small RNAs and I don't think the presence of the ribosomes is necessarily a problem for my downstream goals. I just didn't expect the peak to be so high! I may need to eliminate the possibility that incomplete lysis of my tissue may be responsible for any of the height of the peak at this region and not RNA degradation for some other reason, which I am reasonably sure it is not. If it is not incomplete lysis, and degradation isn't seen near the larger ribosomal subunits (which they are not), then it may be a tissue-specific phenomenon.

I am trying to find a source that explains "Insufficient lysis is indicated by accumulation of small RNA around the 5S rRNA peak."

I appreciate anyone's thoughts!

http://www.nature.com/nprot/journal/...16.090_F2.html
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