title: GSM8729593: A549 cell line, Bacterially supplemented media, replicate 1; Homo sapiens; RNA-Seq
library_construction_protocol: Following incubation, RNA was isolated from LUAD cells using EZNA Total RNA Kit (Omega, Norcross, GA, USA). Libraries were prepared from 1µg RNA using the Universal Plus mRNA-Seq with NuQuant® (Tecan, Switzerland), and Universal Plus UDI S02622 was used to barcode the libraries.  Size, purity, and quantitation of libraries was performed using the Agilent Bioanalyzer (Agilent Technologies, Santa Clara, CA).  Libraries were quantified by Qubit dsDNA HS kit (ThermoFisher, Q33231) and size analysis done with the HS NGS Fragment Kit (Agilent, DNF-474-0500) on the Agilent Fragment Analyzer.  Libraries were diluted to 2nM with Illumina Resuspension Buffer (RSB), equivolume pooled, then further diluted to 750pM with Resuspension buffer with Tween. PhiX positive control was spiked in to a final concentration of 2%. Denaturation and final dilution was done onboard instrument and libraries sequenced on a P2 100 cycle kit (Illumina, 20046811) with 101 sequencing cycles and 2 index reads of 8bp each on the NextSeq 2000 instrument.