GSM9523083: bone marrow; sAA; 1893-A043009; Homo sapiens; RNA-Seq
Quantitative nuclease protection assay (qNPA) of FFPE samples, i.e. extraction free. Library preparation was performed by HTG Molecular Diagnostics, Tucson, AZ, USA using their established protocols. Excess functional DNA nuclease protection probes flanked with universal wing sequences are hybridized to target RNAs. Non-hybridized DNA probes and RNA are digested by S1 nuclease, essentially resulting in a 1:1 ratio of DNA detection probes and initially targeted RNA. DNA protection probes are released after heat denaturalization. Barcodes and adapters were individually added using a 16-cycle PCR reaction. Individual sample purification using AMPure XP beads (Beckman Coulter, Brea, CA, USA) and quantification using  KAPA Library Quantification kit (KAPA Biosystems, Wilmington, MA, USA).