Illumina NovaSeq 6000 paired end sequencing; scRNAseq of mpeg1.1+ cells from lesioned and unlesioned zebrafish larvae spinal cord
scRNAseq of mpeg1.1+ cells from lesioned and unlesioned zebrafish larvae spinal cord
Trunks containing the lesion sites (or equivalent site from unlesioned fish) are collected and kept in PBS on ice.Incubate the trunks (up to 300) in 1 mL of 1X Trypsin-EDTA at 37C for 5-7 mins.Stop dissociation by adding FBS to a final concentration of 5%.Centrifuge at 200g for 7 mins.Discard Sups.Resuspend in 500 uL of PBS.Add the cell suspension into a 40 uM cell strainer.Centrifuge at 200g for 7 mins.Resuspend in the buffer for FACS (PBS 5% FBS) Following cell dissociation and FAC sorting, samples were processed on the 10X Chromium platform using 10X Single Cell 3' v3 chemistry following the manufacturer's guidelines  Following cell dissociation and FAC sorting, samples were processed on the 10X Chromium platform using 10X Single Cell 3' v3 chemistry following the manufacturer's guidelines