A composite immune signature parallels disease progression across T1D subjects (RNA-Seq Cohort 0 WB)

Technical replicate testing was performed to determine the measurement precision of each analyte. Peripheral blood samples were collected in tempus tubes from a healthy control subject with no family history of autoimmunity (n=1) and from subjects with Type 1 diabetes (n=4). Total RNA was isolated independently from 3 replicate aliquots of the same tempus samples and then globin-reduced. RNAseq libraries were prepared from the globin-reduced RNA. Overall design: The replicate testing cohort was used to determine the technical precision of each analyte by calculating the intra-subject coefficient of variation (CV) for three replicate aliquots (n=3) from the same blood draw from 5 subjects. For each analyte, the mean CV across subjects was then calculated. In order to generate the three replicate aliquots, a tempus tube of 9 mL (3 mL blood plus 6 mL tempus solution) was split into three equally sized samples. These samples were processed separately to RNA, then separate globin reduction and library preparation was conducted.