D120_longReads
About 50 ng of total RNA, in a final volume of 9 l, were combined with 1 l of VNP primer (10 M) and 1 l of dNTPs (10 mM), and the mixture was incubated at 65C for 5 minutes. The products obtained in this way were subjected to strand exchange and reverse transcription reactions. Subsequently, cDNA products were labeled with molecular identifiers (barcodes) using the Oxford Nanopore PCR Barcoding Expansion 1-96 kit (code EXP-PBC096), followed by the attachment of adapters from the SQK-DCS109 kit, according to the manufacturer's instructions. Libraries prepared in this way were loaded into a PromethION flow cell (R9.4.1) and sequenced at the Centre de Regulaci Genmica (CRG, Barcelona, Spain).
SUB16008984
D120_longReads