GSM5066621: NeuroD_rep2_overexpression; Ciona intestinalis; RNA-Seq
At the late tailbud stage I, morphologically normal embryos were selected, and at late tailbud II, GFP+ embryos were dissociated as previously described in Cao et al, 2019. The dissociation was performed in duplicate. The cell concentration of each sample was checked by TC20 Automated Cell Counter to ensure it was within 1,000–2,000 cells per microlitre. Single-cell suspensions were loaded onto The Chromium Controller (10x Genomics). For all of the samples, cells were lysed, cDNAs were barcoded and amplified with Chromium Single Cell 3′ Library and Gel Bead Kit v3 (10x Genomics) following the instructions of the manufacturer. Illumina sequencing libraries were prepared from the cDNA samples using the Nextera DNA library prep kit (Illumina). All of the libraries were sequenced on Illumina NovaSeq 6000 with S1 reagent kit (300 cycles, paired-end) following standard Illumina protocols.