Familial Thrombocyotpenia due to a WAC-ANKRD26 Gene Fusion

Human peripheral blood mononuclear cells from patients carrying a WAC-ANKRD26 gene fusion caused by a duplication-inversion-duplication and healthy controls 3 individuals affected with dup-inv-dup, 3 healthy controls. RNA was isolated using a RNeasy Micro kit (Qiagen) according to the manufacturer’s instructions. An on-column DNase digestion was performed before RNA was quantified using a Qubit RNA HS Assay kit (Invitrogen). ~10 ng of RNA was used as input to a modified SMART-seq2 protocol and after reverse transcription, 10 cycles of PCR were used to amplify transcriptome library12,14,15. Quality of whole transcriptome libraries was validated using a High Sensitivity DNA Chip run on a Bioanalyzer 2100 system (Agilent), followed by library preparation using the Nextera XT kit (Illumina) and custom index primers according to the manufacturer’s instructions. Final libraries were quantified using a Qubit dsDNA HS Assay kit (Invitrogen) and a High Sensitivity DNA chip run on a Bioanalyzer 2100 system (Agilent). All libraries were sequenced using Nextseq High Output Cartridge kits and a Nextseq 500 sequencer (Illumina).