RNA-Seq of Homo sapiens: COVID-19,Convalescent,individual035,Visit1
Frozen whole blood samples collected in PAXgene Blood RNA Tubes (PreAnalytiX, #762174) were used for total RNA extraction. RNA integrity and concentration were assessed using a Qubit 2.0 Fluorometer (Thermo Fisher Scientific, #Q32866) and the Qubit RNA HS Assay Kit (Thermo Fisher Scientific, #Q32854). For mRNA enrichment, 200 ng of total RNA per sample was processed using the Dynabeads mRNA Purification Kit (Thermo Fisher Scientific, #01152851), which depletes rRNA and isolates polyadenylated transcripts using oligo(dT) beads. Library preparation was performed using the MGIEasy RNA Directional Library Prep Set (MGI, #MG1000006386), following the manufacturers protocol. Fragment size distribution was checked using 4150 TapeStation (Agilent, #G2992A) with the cDNA D1000 ScreenTape (Agilent, #5067-5582). Final libraries were quantitated using the Qubit 2.0 Fluorometer (Thermo Fisher Scientific, Q32866). RNA sequencing was run on DNBSEQ-T7RS (MGI) platform, generating 150 bp paired-end reads.