title: GSM3498853: small RNA-seq on Dicer KD oocyte; Sus scrofa; miRNA-Seq
library_construction_protocol: Total RNAs from oocytes  were isolated using TRIzol® Reagent (Invitrogen) according to the instructions provided by the manufacturer. RNA concentrations were quantified by measuring absorbance (A260/280ratio) on a NanoDrop Spectrophotometer ND-1000 (NanoDrop). RNA integrity number (RIN) was assessed using an Agilent 2100 Bioanalyzer (Agilent Technologies). Illumina protocols (available on the Illumina website at www.illumina.com/support) were followed to prepare the small RNA libraries from total RNA. Briefly, 3′ and 5′ adapters were ligated to small RNAs (< 200 nt) isolated from total RNAs from oocytes. After adapter ligation, an RT reaction was performed to cDNA synthesis. In order to avoid any bias, the cDNA was PCR amplified by using common primers that were designed against adapter sequences. After cDNA amplification, RCR products were isolated by gel purification and then sequenced by high-throughput deep sequencing using an Illumina® Hiseq2000 sequencer.