SoctosporusAzygotic_19hours_ONT_cDNA_seq
Total RNA was extracted from S. octosporus cells undergoing azygotic meiosis. A total amount of 1ug RNA was prepared for cDNA libraries using protocol provided by Oxford Nanopore Technologies (ONT). Briefly, SuperScript IV First-Strand Synthesis System (Invitrogen) was used for full length mRNA reverse transcription and following cDNA PCR for 14 circles with LongAmp Tag (NEB). The PCR products were then subjected to FFPE DNA repair and end-repair (NEB) steps and following adaptor ligation using T4 DNA ligase (NEB). Agencourt XP beads was used for DNA purification according to ONT protocol. The final cDNA library was added to FLO-MIN109 flowcells and sequenced on the PromethION platform.
SoctosporusAzygotic_19hours_ONT_cDNA_seq