Danio rerio amh mutant RNA-seq
Zebrafish were euthanized in Tricaine. We isolated gonad-containing trunk of the animals by removing the anterior of the fish from just posterior of the pectoral fin and removing the caudal peduncle posterior to the anus. Trunks were individually homogenized in 200ul Trizol. Total RNA was extracted following Amores, 2011. Total RNA was enriched for mRNA using Dynabeads Oligo(dt)25 (ThermoFisher). We constructed indexed, strand-specific cDNA sequencing libraries using the NEXTflex qRNA-seq kit (BIOO Scientific). Library concentrations were quantified using a Qubit fluorometer (Life Technologies), normalized to a concentration of 2.3nM, and multiplexed. Prior to sequencing, we further evaluated the quality of the multiplexed library by quantitative real-time PCR using the Kapa Library Quantification Kit (Kapa Biosystems). One lane of paired-end 100 base pair (bp) sequencing was performed on an Illumina HiSeq 4000.