Rt2 lncrna pcr array human cancer pathway finder

RNA was isolated from cells and exosome samples using the Qiagen miRNeasy kit and the Norgen Biotek Corporation Single Cell RNA Purification Kit (manufacturer’s protocol). For drug treatment, the WRO and WRO-LvR cells were treated with 2 μM KU757 for 48 h. RNA was quantified and its purity validated by NanoDrop. cDNA was synthesized after the elimination of genomic DNA from 1 μg of RNA using the RT² First Strand Kit (Qiagen). For exosomes, cDNA was synthesized after the elimination of genomic DNA from 50 ng of RNA using the RT² PreAMP cDNA Synthesis Kit First (Qiagen; manufacturer’s protocol). cDNA target templates were then pre-amplified using the RT² PreAMP cDNA Synthesis Kit First (Qiagen). Differentially expressed lncRNAs were evaluated using qPCR (Qiagen RT² lncRNA PCR Array Human Cancer Pathway Finder) and analyzed using Qiagen GeneGlobe data analysis.

NCI-H295R cells were treated with 1.0 to 2.0 μM KU758 or 1.0 μM 17-AAG for 24 hours. Total RNA was isolated using the RNeasy Mini Kit (Qiagen, Hilden, Germany) and measured by Nano Drop spectrophotometer (Thermo Fisher Scientific, Waltham, MA). 800 ng of RNA was transcribed to cDNA using the RT² First Strand Kit (Qiagen, Hilden, Germany) and lncRNA expression was determined using SYBR Green quantitative PCR and the RT² lncRNA PCR Array Human Cancer Pathway Finder (Qiagen, Hilden, Germany).