E14_Mprip_dL
For full-length cDNA synthesis, SMARTer PCR cDNA synthesis kit (Clontech) was used according to user manual. Total RNA was DNAse treated either on-column using PureLink DNase (Thermo Scientific 12185010) and PureLink RNA Mini kit (Thermo Scientific 12183020) or with TURBO DNase (Thermo Scientific AM1907). First strand cDNA was synthesized using 700 ng of DNase treated RNA, 3' SMART CDS Primer II A and SMARTer II A TSO. cDNA was diluted 1:5 in water. cDNA was amplified by optimal (18  21) cycles of PCR using Advantage 2 PCR kit (Clontech). cDNA was enriched by capturing with biotinylated xGen Lockdown probes (IDT) and xGen hybridization and wash kit (IDT). Captured cDNA was purified using AMPure XL beads (Beckman Coulter) then amplified using Takara LA Taq DNA polymerase Hot-Start version (Clontech). For nanopore library preparation, captured and amplified cDNA was end-prepped (NEBNext FFPE DNA Repair Mix and NEBNext Ultra II End Repair Kit) and ligated (T4 DNA Ligase  NEB) with sequencing adapters (Nanopore SQK-LSK110). MinION Mk1B device and FLO-FLG001 flow cells were used for sequencing of the libraries.