HPC_donor2_Day3
Total RNA was isolated from 35 HPC samples and 30 MNC samples using the miRNeasy Mini Kit according to the manufacturer's instructions. RNA quantity and purity were assessed using the NanoDrop 8000 spectrophotometer, while RNA integrity was evaluated using the 2100 Bioanalyzer. RNA-seq library preparation and analysis were performed through Illumina reagents. Briefly, cDNA libraries were prepared using the TruSeq Stranded Total RNA Kit (Illumina) and sequenced on the Illumina NextSeq 550 platform. Quality control of raw sequencing reads was performed using FastQC. Reads were aligned to the human reference genome (hg38) using the STAR aligner. SAMtools was used to generate gene count files, and transcript quantification was carried out with featureCounts. Gene expression levels were normalized and reported as TPM (transcripts per million). Batch effects were assessed, and library batch effects were removed from the HPC datasets.