I have a time series experiment and want to compare DE miRNAs between these time points and the Control. I realized that I loos some DE miRNAs upon combining some time points together and conduct the DE analysis, which did not happen when performing the DE analysis on these time points as single. Why this happened ? what is your recommendation ?
Hi Mohamed,
I would say that it looks like it would be something related on which your source tissue is and the miRNA extraction procedure.
I have worked with single time points miRNA colon samples and I succeeded with the extraction kit mirVana miRNA Isolation Kit (Life Technologies). In order to ensure the miRNAs concentration, we add a last precipitation step:
-add acrylamide, sodium acetate and ethanol 100%,
-left it two days at -20ºC,
-purify adding ethanol 80% and centrifugate, twice,
-let the pellet dry under laminar flow cabin for 15 minutes,
-and resuspend it adding 15 uL of RNase free water.
After that, I obtained very good concentrations and very good integrity.
Hope it helped,
Elena
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