GSM5114029: Late L3 larvae (SPLiT-Seq); Caenorhabditis elegans; RNA-Seq
The RNA extraction follows SPLiT-seq protocol SPLiT-seq experiments were performed as described previously (Rosenberg et al., 2018), with a few adaptations according to cell type and population size: (1) 15 μm strainers were used to generate single-cell suspension; (2) fixed worm cells were pellet by centrifuging at 800g for 5 min at 4˚C; (3) cells were counted on a hemocytometer and diluted to 1,000 cells/μL; (4) cells were split into 4 wells for the first round of barcoding (Table S1), each containing barcoded well-specific reverse transcription primers（poly(dT)15 and random hexamer）. Cells were split into 96 wells in round 2 and 48 wells in round 3 well-specific barcoding (Table S1). (5) For tagmentation, the TruePrep DNA Library Prep Kit (Vazyme, #TD501) was used.