title: GSM2348875: LUAD_X0742_T; Homo sapiens; RNA-Seq
library_construction_protocol: Human tumor samples were obtained from patients following written informed consent.Samples were provided as fresh-frozen tissue specimen. Matching normal material was provided in the  form of adjacent non-tumorigenic tissue. All patient material was stored at -80 celsius. After tissue samples treatment, total RNA was extracted from leukocyte by using TRIzol Reagent (Ambion) following the manufacturer’s instructions. Total RNA was treated with RQ1 DNase (promega) to remove DNA. The quality and quantity of the purified RNA were determined by measuring the absorbance at 260 nm/280 nm (A260/A280) using smartspec plus (BioRad). RNA integrity was further verified by 1.5% Agarose gel electrophoresis.  For each sample, 10 μg of total RNA was used for RNA-seq library preparation. Polyadenylated mRNAs were purified and concentrated with oligo (dT)-conjugated magnetic beads (invitrogen) before used for directional RNA-seq library preparation. Purified mRNAs were ion fragmented at 95 ℃ followed by end repair and 5' adaptor ligation. Then mRNA reverse transcription was performed with RT primer harboring 3' adaptor sequence and randomized hexamer. The cDNAs were purified and amplified. And products corresponding to 200-500 bps were purified, quantified and stored at -80 ℃ before sequencing.
attributes.tag: GEO Accession
attributes.value: GSM2348875