Reduction of rRNA during sRNA library construction using a set of blocking oligos, in Drosophila melanogaster

This experiment was designed to improve the proportion of miRNA mapping reads from sRNA library sequencing, by reducing the amount of particularly abundant rRNA fragments. We extracted the sRNA fraction from 50 pooled male D. melanogaster accesory glands and constructed 4 cDNA libraries from the resultant RNA (blocked vs not-blocked with 2 biogical replicates). For the blocked libraries we introduced several blocking oligos, with sequences complentary to the most abundant contaminating rRNA fragments, with the aim of preventing ligation of that rRNA to library adapters. The other libraries (unblocked) were constructed according to standard HD protocol. The libraries were sequenced on a HighSeq2500 platform.