Melanotaenia fluviatilis male brain mRNA sequencing
Total RNA from whole brain was purified using the RNeasy® Lipid Tissue Kit (QIAGEN Pty. Ltd., Australia). On-column DNase treatments were included during purification. RNA integrity was confirmed using the Bioanalzyer 2100 (Agilent). Sequencing was undertaken by a service provider (Australian Genome Research Facility, Melbourne, Australia). Library preparation was performed using Illumina TruSeq reagents and sequencing was conducted using the HiSeq2000 platform. Briefly, RNA quality was determined using a Bioanalyzer 2100 (Agilent) before isolation of polyA+ mRNA, fragmentation, and construction of cDNA libraries using the Illumina TruSeq kit according to the standard protocol. 100-cycle paired-end sequencing was undertaken using two lanes of an Illumina HiSeq2000 instrument, with three libraries multiplexed per lane.