title: Lates calcarifer gonad transcriptome
design: RNA extraction  Total RNA was extracted from the testis of an Asian seabass (Lates calcarifer) individual using Qiagen RNeasy mini kit and followed by Ribosomal RNA depletion using the RiboMinus™ Eukaryote Kit (Invitrogen, P/N A10837-08). Total RNA integrity and the rRNA removal efficiency was confirmed by bioanalyzer analysis.     Whole Transcriptome RNA Library Preparation  The whole transcriptome RNA library was prepared according to the Solid Whole Transcriptome Analysis Kit (ABI, PN 4425680). Briefly, 0.5 μg of rRNA-depleted total RNA was used as starting material, and was randomly fragmented using RNase III. The fragmented RNA sample was next hybridized and ligated with Adaptor Mix A from the kit, and was then reverse transcribed into single strand cDNA, followed by denaturing gel electrophoresis. The size-selected cDNA(100-200nt) was next amplified by in-gel PCR, and barcoded SOLiD 3’ Primer from the SOLiD Transcriptome Multiplexing Kit (ABI, PN 4427046) was used to allow multiplex Solid system sequencing.    SOLiD Sequencing  The cDNA library was clonally amplified onto beads by emulsion PCR using standard protocols ( Applied Biosystems Solid  System Templated Bead Preparation Guide (PN 4407421) ). These beads were then enriched and deposited onto the two octets of a glass slide for sequencing with 50bp reading length.