title: GSM8127891: HAPE patient 005; Homo sapiens; RNA-Seq
library_construction_protocol: Whole blood (2.5 ml) samples from all participants were collected using The PAXgene® Blood RNA Tube (BD, cat. USA), immediately mixed vigorously for 10s and frozen at -80°C until use. The PAXgene Blood RNA Kit (PreAnalytiX) was used to isolate total RNA from whole blood according to the kit instructions. Total RNA quality was assessed using agarose gel electrophoresis and quantified using a NanoDrop2000 spectrophotometer (Thermo Fisher Scientific, USA). The Agilent Bioanalyzer 2100 (Agilent Technologies, USA) was used to verify RNA integrity. The fragmented RNA was used as a template for first strand cDNA synthesis, using random primers (Random Hexamers).The second strand of cDNA was synthesized using DNA polymerase and the double stranded cDNA was purified with magnetic beads (AMPure XP; Beckman Coulter,USA).Then addition of adenlylate 3′ ends, connection sequencing adapters, selection library size, PCR amplification and checking library quality were performed in turn. The whole transcriptome sequencing project requires the construction of two sequencing libraries simultaneously, including a strand-specific library for rRNA removal and a small RNA library. The constructed library was sequenced using Illumina high-throughput sequencing platform according to standard protocol (Genesky Biotechnologies Inc., Shanghai, China). Detailed experimental procedures are provided in the Supplementary Material. This study obtained the expression landscape of mRNAs, miRNAs, circRNAs, and lncRNAs from the same samples in the discovery set and conducted subsequent bioinformatics analysis.The cDNA library was sequenced on an Illumina NovaSeq 6000 sequencing platform to generate read pairs of 150 bp length.