RNA-Seq of  zebrafish otop1 gene knock-in mus musculus: adult male DRG
After extracting total RNA from the sample, magnetic beads with Oligo(dT) were used to enrich eukaryotic mRNA . MRNA was fragmented into short fragments by adding fragmentation buffer. The first cDNA strand was synthesized with six-base random primer using mRNA as template. Then buffer, dNTPs, RNase H, and DNA polymerase I were added to synthesize A second cDNA strand, which was purified by QiaQuick PCR kit and eluted with EB buffer for end repair, poly (A) addition, and sequencing adaptor attachment. Then, the fragment size was selected by agarose gel electrophoresis, and the final PCR amplification was performed. The constructed sequencing library was sequenced by Illumina HiSeq.