GSM7898625: 10mT_fractured_2; Danio rerio; RNA-Seq
Total RNAs were extracted from scales using RNeasy Mini Kit (QIAGEN). 3' RNA-seq was performed according to the Lasy-Seq Ver 1.1 method (Kamitani et al. 2019; Kashima et al. 2022). Total RNA was extracted from zebrafish scales using RNeasy Mini Kit (QIAGEN). Reverse transcription (RT) was performed using Super Script IV (Thermo Fisher Scientific) and an RT primer containing oligo-dT, the index sequence, and 9-bases unique molecular identifiers (UMIs). RNA/cDNA hybrids were purified using AMpure XP beads (Beckman Coulter), and second-strand DNA was synthesized using DNA polymerase I (Enzymatics). dsDNAs were fragmented using WGS Fragmentation Mix (Enzymatics) and ligated into customized adapters with WGS Ligase (Enzymatics). Adapter-ligated dsDNAs were amplified with KAPA HiFi ReadyMix (Nippon Genetics) and SE PCR primers. Next-generation sequencing of cDNA libraries was performed by Novogene using an Illumina NovaSeq X (illumina).