title: MinION sequencing
library_construction_protocol: All strains were cultured at 20°C on nematode growth medium (NGM) plates seeded with E. coli HB101 as a food source. Wild-type or tent-5(tm3504) young adult males were hand-picked from the male-enriched plates and transferred to an Eppendorf tube containing 10 μl of 50 mM NaCl. The worm pellet was then resuspended in 200 μl of TRI Reagent (Sigma-Aldrich), vortexed for 15 min at room temperature, and stored at -80°C before RNA isolation. TRIzol samples from three different pickings were combined just before RNA isolation, resulting in approximately 300 male worms per sample. The wild-type hermaphrodite population was age-synchronized by bleaching gravid adults and seeding isolated embryos on plates. Worms were grown until they reached the young adult stage, then collected and washed three times using 50 mM NaCl. The worm pellet was resuspended in 1 ml of TRI Reagent (Sigma-Aldrich), vortexed for 15 min at room temperature, and stored at -80°C. RNA was isolated from TRIzol according to the manufacturer's instructions. The samples were additionally purified with KAPA Pure magnetic beads (RNA to beads ratio: 1:3 v/v), and RNA quality was assessed using the Agilent TapeStation system. Sequencing libraries were prepared using the DRS Kit (Oxford Nanopore Technologies, SQK- RNA002) according to the manufacturer's protocol. In each library, mRNA isolated from male or hermaphrodite worms (0,5 - 1,5 μg depending on the amount of isolated RNA) was mixed with 2 ng of in vitro transcribed poly(A) standards nad mRNA isolated from mice. 