Fig. 1

Blood transcriptomics workflow from sample collection to data report. (1) Blood samples from all species (human and model animals) were collected on PAXgene tubes. The PAXgene buffer stabilized samples before extraction. After the first step-by-step freezing, samples were frozen at -80 °C until extraction. Samples came from 6 donors for human, rabbit, and mouse, and 4 donors for NHP in triplicate. (2) Total RNA was manually extracted with Maxwell HT Simply RNA kit custom (#AX2420, Promega). RNA was then processed using the RNA Clean and Concentrator kit, including an additional DNase-I treatment (#R1013, ver.2.2.1, Zymo Research). (3) Total RNA libraries were prepared using the Zymo-SeqRiboFree Total RNA Library Kit (#R3003, Ver.1.04, Zymo Research) which integrates the depletion of globin mRNA and rRNA. The conditions of the library preparation were adjusted according to the associated species and the extraction yields. Libraries were sequenced on the NextSeq500 system (Illumina). (4) Quality control of the data was performed using an in-house pipeline that includes four main stages: (i) quality assessment, (ii) read mapping, (iii) transcript quantification and (iv) filtering and normalization. (5) The pipeline generated a final report assembling all the necessary plots to evaluate the quality of the data