Instrument: Illumina HiSeq 4000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: Total RNA was isolated using STAT-60 (AMS biotechnology, CS-111)/chloroform (200 uL; Sigma, Cat 650471)/isopropanol (500 uL; Sigma, cat 33539)/75% ethanol (1ml) according to previously published protocol (PMID: 26679101). Samples were stored in aliquot at – 80 °C prior to use. All reagents, plastic ware, and supplies used were sterile, nuclease free, and of molecular biology grade. RNA purity (A260/A280>1.80) and concentration were determined using nanodrop spectrophotometer (Thermo Fisher scientific, Delaware USA). RNA integrity was studied using the 2100 Bioanalyzer System (Agilent) and RNA 6000 Nano Kit. A RNA Integrity Number (RIN) of 8 was considered the lowest cut-off for RNA Sequencing. Sequencing library were constructed using Illumina mRNA library prep kit.