alias: GSM3595901
title: GSM3595901: S1318N; Homo sapiens; RNA-Seq
library_construction_protocol: RNA of all fresh frozen tumors and corresponding normal gastric mucosa were extracted using the TRIzol® Reagent in accordance with the protocol provided by the manufacturer (User Manual TRIzol® Reagent, Invitrogen). In brief, 1 mL of TRIzol was added to every 4 mm3 of fresh tissue, followed by the addition of 200 μL of chloroform in each tube. The RNA in the aqueous phase was precipitated with 0.5 ml of isopropanol and washed with 1 ml of 75% ethanol. The resulting RNA pellet was then resuspended in 50 μL of DEPC-treated water and quantified using a NanoDropTM 1000 (Thermo Scientific) to determine the sample concentration and purity. The use of degraded RNA can result in low yield, overrepresentation of the 5' ends of the RNA molecules, or failure of the whole transcriptome sequencing library preparation. Total RNA integrity was evaluated following isolation using an Agilent Technologies 2100 Bioanalyzer, and samples with an RNA Integrity Number greater than 5.0. RNA was quantified with rRNA ratio≧1.5, and amount of RNA≧1.2 µg. The Illumina TruSeq RNA Library Preparation Kit v2 was used for the mRNA-focused sequencing library
identifiers.namespace: GEO
identifiers.id: GSM3595901