GSM9105535: 3dpf_fibroblasts_sort_500_cells_injury_sample3; Danio rerio; RNA-Seq
The fibroblasts were isolated by FACS sorting and cDNA library was constructed by Smartseq2 protocol. In this experiment, we have conducted acetic acid corrosion in four trunk sites in 2 dpf embryos,cut the trunk region at 3 dpf, dissociated the embryos into single cell suspension and performed FACS sorting of dsRed positive fibroblasts. In each biological replicate, around 30 embryos were harvested and 500 fibroblsts were sorted into lysis buffer. Smartseq2 protocol was utilized for double strand cDNA library construction. After acquiring the cDNA from sorted fibroblasts, 0.5 ng cDNA input was tagmented using the TruePrep DNA Library Prep Kit V2 for Illumina (TD503, Vazyme). Following tagmentation, the samples were amplified using the TruePrep Index Kit V2 for Illumina (TD202, Vazyme) for 12 cycles and subsequently purified using Ampure XP beads (A63881, Beckman Coulter). The quality of the libraries was evaluated using the Agilent Fragment Analyzer System and then sent to Novogene for Illumina Hiseq X Ten 150-bp paired-end sequencing, with an average sequencing depth of 8 × 10^6 raw reads per sample.