I want to know which reference gene is best for the normalization of micro RNAs in qRT-PCR. My research plant is coming under the family Leguminosae. Is there any plant or family-specific reference genes?
It's hard to give a specific answer here but, outside of the usual Go-Tos (GAPDH, 18S rRNA, bActin, etc.,), I think the best reference genes would be the ones that represent what you are studying the leguminosae family for. Are you investigating the impact of stressors such as dryness/drought, osmotic/salinity, and temperature (hot/cold)? And what are you focusing on, root? Stem? Leaves? These would determine the legume-specific ref genes that you would want to use. Perhaps focus on the miRNA pathway regulators. What has also helped me with my work is including ref genes that have low, medium, and high expression. That way, no matter the expression level of my target gene(s), it is all covered and accurately normalized. You can also consider the miRNAs that regulate the miRNA processing pathway (i.e., AGO), or miRNAs that regulate the stressors I mentioned earlier.