title: GSM8281735: HUVEC, control, biological replicate 2 (C2); Homo sapiens; RNA-Seq
library_name: GSM8281735
library_construction_protocol: HUVEC cells were grown in dishes, either left untreated or subjected to inflammatory stressor TNF alpha for 6hrs or febrile-mimicking heat stress for 2hrs and cryopreserved in 1ml of freezing media containing more than 0.4x10^6 cells. The experiments were done in biological triplicate. RNA-seq was done by Diagenode Inc. Specifically, nine cryopreserved samples were used for total RNA extraction using the Qiagen RNeasy mini kit (Qiagen Cat #74104). The RNA-seq experiment has been conducted by RNA-seq services (Diagenode Cat# G02030000). RNAs have been firstly quantified using Qubit™ RNA HS Assay Kit RNA HS Assay Kit (Thermo Fisher Scientific, Cat# Q32852). The RNA integrity was assessed using a Bioanalyzer, RNA 6000 Pico kit (Agilent  Cat# 5067-1513,) on a 2100 Bioanalyzer system (Agilent). The samples with RINs above 7 were used for RNA-seq. Library preparation has been done with 250ng of input RNA using NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB #E7490) followed by NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (NEB #E7760) and NEBNext® Multiplex Oligos for Illumina® Index Primers Set 1 (NEB# E6440). The generated DNA libraries were purified using Agencourt® AMPure® XP (Beckman Coulter). Purified libraries were quantified, and their size was assessed with QIAxcel (Qiagen). The library pool was then sequenced on NovaSeq paired-end 50bp.