GSM2407513: SiChe-1 4H Hypoxia; Homo sapiens; RNA-Seq
HCT116 cell line was transfected with siChe-1 or siRNA negative control using Lipofectamine 3000 (Life Technologies), total RNA was extracted from cells using TRIzol reagent (Life Technologies) in accordance with the manufacturer's instructions. Quantity and integrity of the extracted RNA were assessed by NanoDrop Spectrophotometer (NanoDrop Technologies) and by Agilent 2100 Bioanalyzer (Agilent Technologies), respectively. RNA libraries for sequencing were generated according to the standard Illumina TruSeq RNA sample preparation protocol using 2ug total RNA as starting material. The resulting library was controlled qualitatively with the High Sensitivity DNA Kit (Agilent Technologies, Santa Clara, CA, USA) and quantitatively with real-time analysis employing a SYBR Green qPCR protocol with specific primers complementary to adapter sequences. Therefore, only the adapter-ligated fragments that are appropriate for sequencing were quantified. Based on the qPCR quantification, libraries were normalized to 1 nM and denatured by using 0.1N NaOH. Cluster amplification of denatured templates was carried out according to manufacturer protocol (Illumina, Inc., San Diego, CA, USA). Sequencing was performed on a Genome Analyzer IIx (Illumina) in paired-end mode, sequencing from each side 51 bp.
GEO Accession
GSM2407513