title: GSM8170412: ONT_cDNAseq_LV3691_TASOR_CRISPRi; Homo sapiens; RNA-Seq
library_construction_protocol: 1.5 μg total RNA was isolated as above for each of the four samples tested (Control, MORC2-CRISPRi, TASOR-CRISPRi and DNMT1-KO hNPCs, 10 days post-transduction in each case). Ribosomal RNA was depleted using the RiboMinus Eukaryote System v2 (ThermoFisher). 5 ng of the resulting RNA was used to make barcoded cDNA libraries (2 barcodes per sample) using the cDNA-PCR kit SQK-PCB-111-24 (Oxford Nanopore Technologies), which incorporates a strand-switching step to enrich for full-length transcripts. Fifteen PCR cycles were used with 7 min extension time. Inspection of the final cDNA product by capillary electrophoresis confirmed a peak at around 2-2.5 kb, approximately the size of an average human mRNA. The libraries were sequenced with a FLO-PRO002 PromethION Flow Cell R9 Version on a PromethION (Oxford Nanopore Technologies) at SciLifeLab