title: GSM4594784: channel catfish 8dpf_rep2; Ictalurus punctatus; RNA-Seq
library_construction_protocol: RNA extraction was conducted using the Qiagen RNeasy Plus Kit following the manufacture protocols. The RNA integrity was checked with 4200 TapeStation (Agilent Technologies, Palo Alto, CA, USA). Ribosomal RNA depletion was conducted using Ribozero rRNA Removal Kit (Illumina, San Diego, CA, USA). RNA sequencing library preparation was performed using NEBNext Ultra RNA Library Prep Kit for Illumina (NEB, Ipswich, MA, USA). Enriched RNAs were fragmented for 15 minutes at 94 °C. The first strand and second strand cDNA were subsequently synthesized. The cDNA fragments were end-repaired and adenylated at 3'ends, and a universal adapter was ligated to cDNA fragments, followed by index addition and library enrichment with limited cycle PCR. Sequencing libraries were validated using the Agilent Tapestation 4200 (Agilent Technologies, Palo Alto, CA, USA) and quantified by using Qubit 2.0 Fluorometer (Invitrogen, Carlsbad, CA).