title: GSM8763338: HeLa, RNA-seq, replicate 1; Homo sapiens; RNA-Seq
library_construction_protocol: Total cellular RNA was isolated using TRI Reagent (Sigma). DNA contamination was removed by digestion with TURBO DNase (Thermo Scientific). The RNA was then purified by phenol:chloroform extraction, precipitated with ethanol, suspended in water, and measured in a NanoDrop ND2000 spectrophotometer (Thermo Scientific). A total of 2 mg of purified total RNA was ribodepleted in cytoplasmic rRNA using the Ribo-Zero rRNA Removal Kit (Illumina) according to the manufacturer's instructions. The KAPA RNA HyperPrep Kit (Kapa Biosciences) was used to prepare the strand-specific libraries according to the manufacturer's instructions. To estimate the number of cycles for library amplification, the KAPA Library Quantification Kit (Kapa Biosciences) was used according to the manufacturer's instructions. The final amplification products (cDNA libraries), in addition to purification with the KAPA RNA HyperPrep Kit, were purified from short fragments on AMPure XP (Beckman Coulter) and then analyzed on the 2100 Bioanalyzer using the High Sensitivity DNA Kit (Agilent). cDNA libraries were prepared in three biological replicates.