Depletion of erythropoietic miR-486-5p and miR-451a improves detectability of rare microRNAs in peripheral blood-derived small RNA sequencing libraries

Erythroid-specific miR-451a and miR-486-5p are two dominant microRNAs (miRNAs) in human peripheral blood-derived small RNA sequencing libraries. Their overabundance reduces diversity as well as complexity of libraries after PCR amplification and consequently causes negative effects such as inaccurate detectability and quantification of other low abundant miRNAs. Here we present a cost-effective and feasible hybridization-based method to deplete these two erythropoietic miRNAs from blood-derived RNA samples Overall design: In order to test the efficiency of our blocking oligos targeting the erythropoietic miRNAs, for each replicate sample, we generated unblocked (standard) and blocked (modified) libraries using both TruSeq and NEXTflex protocols. With this design, we generated 8 paired libraries for TruSeq and 5 paired libraries for NEXTflex using whole blood total RNA as an input.