GSM9786009: Leucoraja erinacea spinal cord bulk RNA-seq, biological replicate 1; Leucoraja erinaceus; RNA-Seq
Spinal cord and dorsal root ganglia (DRG) were dissected separately from stage 32 Leucoraja erinacea embryos and stored in TRIzol reagent (Invitrogen) at -80°C. Samples were homogenized by trituration through 18-gauge (20x) and 25-gauge (10x) needles. Following addition of 100 µL chloroform, samples were incubated on ice for 5 minutes and centrifuged at 12,000 rpm for 15 minutes at 4°C. The aqueous layer was transferred to a new tube, mixed with 250 µL isopropanol, incubated on ice for 10 minutes, and centrifuged at 12,000 rpm for 10 minutes at 4°C. RNA pellets were washed twice with 500 µL 75% ethanol (7,000 rpm, 5 minutes, 4°C), air-dried, and resuspended in 25 µL EB buffer. Samples were further purified using the RNA MinElute Cleanup Kit (Qiagen) to remove DNA and small RNA contamination. Stranded RNA-seq libraries were prepared with polyA selection by the NYU Langone Genome Technology Center (RRID: SCR_017929). Libraries were sequenced on an Illumina NovaSeq 6000 using an SP flow cell (300 cycles, v1.5 chemistry) with paired-end reads.