GSM4154890: NG 6; Homo sapiens; ncRNA-Seq
Before the sequencing experiment, we evaluated the integrity and quantity of each RNA sample using and Nano drop TM instrument. Total RNA from patients' sera were isolated with TRIzol LS Reagent (Life Technologies, USA), following the manufacturer's protocols. The concentration and purity of RNA can be determined by the NanoDrop®ND-1000. The ratio of A260 to A280 values is a measurement of RNA purity, and it should fall in the range of 1.8 to 2.1. Final RNA preparations were resuspended in RNase-free water and stored at -70°C. Total RNA of each sample was sequentially ligated to 3' and 5' small RNA  adapters. cDNA was then synthesized and amplified using Illumina's  proprietary RT primers and amplification primers. Subsequently, ~134-160 bp  PCR amplified fragments were extracted and purified from the PAGE gel. And  finally, the completed libraries were quantified by Agilent 2100 Bioanalyzer.