Channel catfish RNA-seq
Total RNA was isolated using the RNeasy plus Mini Kit (Qiagen, Valencia, CA, USA) with DNase I (Invitrogen, USA) treatment following the manufacturer's protocol. Total RNA was amplified using NuGEN Technologies Ovation RNA-seq product. The produced double-stranded cDNA was subsequently used as the input to the Illumina library preparation protocol starting with the standard end-repair step. The end-repaired DNA with a single A-base overhang is ligated to the adaptors in a standard ligation reaction using T4 DNA ligase and adaptor, depending on the DNA yield following purification after the addition of the A-base. Following ligation, the samples were purified and subjected to size selection via gel electrophoresis to isolate 350 bp fragments for ligation-mediated PCR (LM-PCR). LM-PCR were used to amplify the ligated material in preparation for cluster generation.