GSM8863177: sorted mCherry+ tenocytes from 36 hpf Tg(scxa:mCherry) zebrafish embryos replicate 4; Danio rerio; RNA-Seq
36 hpf and 48 hpf Tg(scxa:mCherry) zebrafish embryos (labeling tendon fibroblasts/tenocytes) were dissociated with Collagenase IV (Gibco, 17104019) at a concentration of 6.24 mg/ml without trypsin addition at a temperature of 28C for roughly 40 min, homogenizing every 5 min with a P1000 pipette. Cells were filtered through a 40um filter (Pluriselect-usa, 43-10040-50). Dissociated cell suspensions were sorted on a Bio-Rad FACS Aria II cell sorter. mCherry-positive cells were gated and sorted for those expressing at high levels. RNEasy Micro Kit (Qiagen, 74004) was used for RNA extraction of cell lysates from FAC-sorted cells, and the Smart-seq2 protocol was utilized for cDNA library construction. Libraries were sequenced using a Hi-seq 4000 sequencer (Illumina) at a read depth of ~35M reads per replicate.