Fig. 1

Experimental design of an RNA-seq based diagnostic study. First, individuals suspected of a Mendelian disorder are recruited for DNA sequencing. In addition, patient biopsy material is collected during the routine medical examination and prepared for RNA extraction. The sample preparation process can take from hours for biopsies to weeks for establishing a cell culture. RNA sequencing is then performed followed by alignment and quality control. The generated data go through DROP which consists of quality control steps and detection of aberrant RNA expression events. The results are then interpreted by sample, including the association of aberrant RNA expression events with rare variant(s) and the function of affected genes with the patient phenotype, which can lead to new diagnoses or candidates. Experience-based estimated durations are provided for each step