Heat-shock influence on gene expression in HEK293T human cells.

RNA-Seq procedure was used for analysis of expression in HEK293T cells. Heat shock treatments was performed for 20 min at 43°C. Then heat shock treated cells were incubated for 6 h at 37°C and replicates were separately lysed with Trisol reagent. Total RNA was extracted from cell lysed with Trisol reagent and PureLink RNA Micro Kit (Invitrogen) according to manufacture instruction. RNA Quality was checked with BioAnalyser and RNA 6000 Nano Kit(Agilent). Poly(A)+ RNA was isolated with Dynabeads® mRNA Purification Kit (Ambion). Illumina library was prepared from poly(A)+RNA with NEBNextNEBNext® Ultra™II RNA Library Prep Kit for Illumina® (NEB) according to manual. Sequencing was performed on HiSeq1500 with 50 bp read length. Analysis of HEK293T gene expression levels by RNA-Seq.