GSM3932592: SI02; Mus musculus; RNA-Seq
Animals were perfused with aCSF supplemented with ion channel inhibitors. Lumbar spinal cord segments were dissected and dissociated in aCSF supplemented with 2mg/mL Collagenase + 2mg/mL Protease from streptomyces griseus. A percoll gradient was used to isolate cells from debris. Cells were then methanol fixed and stored in -80 before being rehydrated for Drop-seq. Drop-seq was carried out as described (Loo et al 2019). RNAs were captured with Chemgenes oligodT barcoded beads (Macosko2015). Reverse transcription was performed using Maxima H Minus Reverse Transcriptase (ThermoFisher; EP0751). Libraries were amplified with KAPA Hi-Fi master mix and tagged using a Nextera XT DNA sample preparation kit (N701-N707, N711, N712, N714-N726) Single-cell RNA seq (Drop-seq); Pooled libraries