GSM3177174: wildtype EPAS1+/+ 786-O cells, replicate 1; Homo sapiens; RNA-Seq
Total RNA was extracted from adherent 786-O cells using the RNeasy Mini Kit (Qiagen Inc.) according to manufacturer's instructions. RNA sequencing library was prepared using NEBNext Ultra RNA Library Prep Kit following manufacturer's recommendations. Briefly, mRNAs were first enriched with Oligod(T) beads and fragmented for 15 minutes at 94 °C. First strand and second strand cDNA were subsequently synthesized, end repaired and adenylated at 3'ends. Universal adapter was ligated to cDNA fragments, followed by index addition and library enrichment with limited cycle PCR. Sequencing libraries were validated using the Agilent Tapestation 4200, and quantified using Qubit 2.0 Fluorometer as well as by quantitative PCR. The sequencing libraries were multiplexed and clustered on one lane of a flowcell. After clustering, the flowcell was loaded on the Illumina HiSeq instrument according to manufacturer's instructions.  The samples were sequenced using a 2x150 Paired End (PE) configuration.  Image analysis and base calling were conducted by the HiSeq Control Software (HCS). Raw sequence data (.bcl files) generated from Illumina HiSeq was converted into fastq files and de-multiplexed using Illumina's bcl2fastq 2.17 software. One mis-match was allowed for index sequence identification.  
GEO Accession
GSM3177174