small RNA-seq of from Drosophila simulans ovaries, control condition
Small RNAs were isolated using anion exchange chromatography                 extraction, and then 18-40 nt were size selected on gel. Small RNA-Seq librairies were generated using                 TruSeq Small RNA Library Prep Kit (Illumina, San Diego, CA), according to manufacturer's instructions.                 The protocol takes advantage of the natural structure common to most microRNA molecules that have a 3'                 hydroxyl group resulting from enzymatic cleavage by Dicer or other RNA processing enzymes. Briefly, in                 the first step, RNA adapters were sequentially ligated to each end of the RNA, first the 3' RNA adapter                 that is specifically modified to target microRNAs and other small RNAs, then the 5' RNA adapter. Small                 RNA ligated with 3' and 5' adapters were reverse transcribed and PCR amplified (30 sec at 98oC; [10 sec                 at 98oC, 30 sec at 60oC, 15 sec at 72oC] x 13 cycles; 10 min at 72oC) to create cDNA constructs.                 Amplified cDNA constructs of 140 to 160 nt were selectively isolated by acrylamide gel purification                 followed by ethanol precipitation. The final cDNA libraries were checked for quality and quantified                 using capillary electrophoresis.
SUB7716124
dssj9_ovaries_control_rep2