title: ZZ_5d_biological replicate 3
design: Total RNA was extracted from gonads of ZZ and ZW fishes using the Trizol Reagent (Invitrogen, Carlsbad, CA) according to the manufacturers instruction, and purified using DNaseI to eliminate genomic DNA contamination. RNA degradation and contamination were monitored on 1% agarose gels and a Nanodrop spectrophotometer. Illumina sequencing was carried out at Novogene Bioinformatics Technology Co., Ltd., in Beijing, China. Sequencing libraries were constructed using the NEBNext Ultra RNA Library Prep Kit for Illumina (NEB, USA), according to the manufacturers protocol. Index codes were added to attribute sequences to each sample. The prepared libraries were sequenced on an Illumina Hiseq 2500 platform, and 150bp paired-end reads were generated.