RT-qPCR Analysis of Exosomal miRNAs has become one of the main talks among researchers, thanks to its contribution to unraveling many cell-to-cell communication mechanisms.
Exosomes are small (30–150 nm) vesicles secreted by cells, performing as key regulators in cell-to-cell communication in various conditions. The molecular constituents of exosomes, especially exosomal microRNAs (miRNAs), have become the focus of many studies.
These studies have shown that exosomes can transport mRNA and miRNA between cells and that the transferred RNA is functional in its new location. Being secreted, exosomes occur naturally in body fluids, making plasma and serum exosome miRNA profiling a potential tool in the diagnosis of different diseases, and in the discovery of new Biomarkers.
Check this article where plasma and exosome miRNA expression levels were used as suggested prognostic marker.
Findings like these can be further implemented also in basic research of tumor cells and their interactions with their microenvironment
If preparing buffers, use water that is nuclease-free or treated with DEPC. DEPC (diethylpyrocarbonate) reacts with histidine residues of proteins and will inactivate RNases. However, it can also react with RNA, so it needs to be removed by heat treatment before the solution is used (DEPC breaks down to CO2 and ethanol).
Set up designated RNA work areas that have low traffic and are away from air vents
Protect your samples with reagents such as RNAlater Stabilization Solution and store the RNA in proper conditions:RNA is generally stable at -80° C for up to a year without degradation in RNase-free H2O with 0.1 mM EDTA or TE buffer (10 mM Tris, 1mM EDTA).
RNA solubilized in formamide may be stored at -20°C without degradation for at least one year
For long term storage, RNA samples may also be stored at -20°C as ethanol precipitates.
More information regarding RNA Preservation and Stabilization can be found in this article.