GSM8694835: Cre_NT, PE250; Chiloscyllium plagiosum; ChIP-Seq
Two weeks after the final vaccination, the sharks were sacrificed using humanitarian execution methods and the spleen was then carefully isolated and stored at -80 °C for future study.Total RNA from the spleen was isolated using TRIzol (Thermo Fisher Scientific) according to the manual. cDNA synthesis was carried out using the SuperScript III First-Strand Synthesis System (Thermo Fisher Scientific). VNAR libraries for next-generation sequencing were prepared, utilizing the primers provided.The PCR products ranging from 300 to 500 bp were recovered. In our assay, the PCR cycles ranged between 25 and 30 cycles. Sequencing libraries were prepared with the NEBNext® Ultra™ II DNA Library Prep Kit (NEB, USA) as the manufacturer's recommendations. Indexes were added to attribute sequences to each sample.