small RNA deep-sequencing with oxidation/ß-elimination with age in Drosophila

Drosophila miRNAs show distinct change in isoform distribution pattern with age. Some miRNAs show accumulation of the short isoforms, while other miRNAs show the accumulation of the long isoforms with age. The increase of the long isoforms of some miRNAs reflects increased 2''-O-methylated miRNA isoforms with age. This raised a question on whether additional miRNAs show increased 2''-O-methylated miRNA isoforms with age. To investigate change in 2''-O-methylated small RNA population with age, we performed small RNA deep-sequencing of oxidized or non-oxidized small RNAs at 3d and 30d in Drosophila. This analysis revealed change in global pattern of 2''-O-methylated miRNA isoforms with age. Overall design: 3d and 30d wild-type male flies were collected. Total RNA was prepared with Trizol reagent, followed by oxidation/ß-elimination assay or the control treatment (no-oxidation). Small RNA libraries were prepared using Illumina''s TruSeq small RNA sample preparation kit (#RS-200-0012, Illumina, Inc. San Diego, CA), following the manufacturer''s protocol. The libraries were multiplexed and sequenced on HiSeq2000 platform (Illumina).