title: GSM8487971: CxCC_79_HPV52; Homo sapiens; RNA-Seq
library_name: GSM8487971
library_construction_protocol: Total RNA was extracted using the TRIzol method, and prepared using Rneasy Mini and Micro Kit following manufacturer's instructions. Library preparation was performed with 10ug of total RNA. Sample quality was assessed with Agilent bioanalyzer or tapestation. Ribosomal RNA was removed by poly-A selection using Oligo-dT beads (mRNA Direct kit, Life Technologies). mRNA was then fragmented in buffer containing 40mM Tris Acetate pH 8.2, 100mM Potassium Acetate and 30mM Magnesium Acetate and heating to 94 degrees for 150 seconds. mRNA was reverse transcribed to yield cDNA using SuperScript III RT enzyme (Life Technologies) and random hexamers. A second strand reaction was performed to yield ds-cDNA. cDNA was blunt ended, had an A base added to the 3' ends, and then had Illumina sequencing adapters ligated to the ends. Ligated fragments were then amplified for 12 cycles using primers incorporating unique index tags.