title: GSM7778666: psmb1 mutants, 72hpf, biological replicate 3, cranial mylz2+ cells; Danio rerio; RNA-Seq
library_construction_protocol: 20-25 heads per sample were dissected from 72hpf larvae. Dissociation was carried out using 25ug/ml liberase (Sigma-Aldrich 05401119001), and samples were incubated on a 600rpm shaker at 37C for 30 minutes. To encourage dissociation, samples were pipetted 20-25 times every 8-10 minutes. Samples were then passed through a 40 μm filter. Cells were sorted into Buffer RLT with 1% β-mercaptoethanol and RNA was isolated using the Qiagen RNeasy MicroKit (Qiagen 74004). For RNA-seq on sox10:kaede+ cells, 7000 cells were sorted per sample. For RNA-seq on mylz2+ cells, 1500 cells were sorted per sample. The SMART-Seq HT kit was used for full-length cDNA synthesis and amplification (Takara, San Jose, CA, USA), and the Illumina Nextera XT kit (Illumina, San Diego, CA, USA) was used for sequencing library preparation.