title: GSM5944745: Differentiated HL-60 neutrophil-like cells processed by capsaicin for 8 hours rep-2; Homo sapiens; RNA-Seq
library_construction_protocol: Total cell RNA was extracted using Trizol (Ambion, USA), every 5*10^6 cells for 1ml Trizol. Then add one-fifth of the total volume of chloroform, shake vigorously for 15 seconds, stand at room temperature for 3 minutes, and rotate at 12000 rpm for 15 minutes at 4°C. Then the upper water phase was transferred to a new centrifuge tube with equal volume of isopropyl alcohol and bottom-up to mix, and incubate at room temperature for 10 minutes. Centrifuged at 12000 rpm for 10 minutes at 4℃. Discard the up and added 1ml 75% alcohol diluted with RNase free water. RNA degradation and contamination was monitored on 1% agarose gels. RNA purity was checked using the Nanophotometer ® spectrophotometer (IMPLEN, CA, USA). RNA integrity was assessed using the RNA Nano 6000 Assay Kit of the Bioanalyzer 2100 system (Agilent Technologies, CA, USA). Sequencing libraries were generated using NEBNext® UltraTM RNA Library Prep Kit for Illumina® (NEB, USA) according to the manufacturer's recommendations and index codes were added to the attribute sequences for each sample