We performed next generation sequencing of miRNAs and then did in situ hybridisation of some of the identified miRNAs. The relative in situ signal strength and read number between some miRNAs seems to be correlated, but not in other cases. I.e. if the read number was, let's say, 60 miRNAX and 20 for miRNAY than signal was stronger for X than Y, but when read number was 30000 for miRNAZ and 5 for miRNAK, than the signal was stronger for K than for Z.
I would appreciate any possible explanation for this!
pl see the scholarly article in link
https://scholar.google.co.in/scholar?q=How+miRNA+NGS+abundance+correlates+with+signal+intensity+in+in+situ+hybridisation%3F&hl=en&as_sdt=0&as_vis=1&oi=scholart&sa=X&ved=0ahUKEwitl9jtkbnWAhUILY8KHRELCBYQgQMIIzAA
Ferenc - interesting question. Have you checked the secondary and tertiary structure of the miRNAs? I know that some have very extensive internal complementation and that harsh conditions are needed to reduce them to their primary structure. Hope this helps.
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