Instrument: Illumina HiSeq 4000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: Cervical and lumbar segments were dissected from 2 neonates at P0 in aCSF, and cell dissociation was conducted using Papain following the manufacturer's instruction (Worthington Biochemical) with 5% Trehalose, 50uM APV, and 800uM KA added during the dissociation process (Saxena et al., 2012). Via FACS (FACSDIVA, BD), 2800 cervical cells and 2800 lumbar cells were collected into 500ul of FACS buffer (Sternfeld et al., 201). Cell suspension was centrifuged and resuspended in 60ul of FACS buffer, 33.4ul of which was loaded onto the 10X Chromium Controller using Chromium Single Cell 3' v2 reagents. Sequencing library was prepared following the manufacturer's instructions (10X Genomics), with 12 cycles used for cDNA amplification and 11 cycles for library amplification.