title: Ion Torrent S5 sequencing; Global gene expression profiles of 10 gastric cancer (GC) samples and 10 gastric normal adjacent tissue (gNAT) samples
design: Global gene expression profiles of 10 gastric cancer (GC) samples and 10 gastric normal adjacent tissue (gNAT) samples
library_construction_protocol: All samples were placed in RNAlater RNA Stabilization Reagent (QIAGEN, Germany) and stored at -80ºC before RNA extraction.  Total RNA was extracted with TRIzol (Ambion, USA) reagent according to the manufacturer's instructions. A barcoded cDNA library was first generated with SuperScript® VILO™ cDNA Synthesis kit from 10 ng of total RNA. Then cDNA amplified using Ion AmpliSeq™ Transcriptome Human Gene Expression Kit (ThermoFisher) to accurately maintain expression levels of all targeted genes. The average size of each amplicon is ~150 bp. Amplified cDNA Libraries evaluated for quality and quantified using Agilent Bioanalyzer High sensitivity chip. Libraries were then diluted to 100pM and pooled equally, with eight individual samples per pool. Pooled libraries amplified and enriched by using IonChef Instrument (ThermoFisher) according to manufacturer instructions.