title: Lates calcarifer gonad transcriptome
design: RNA extraction:  Total RNA was extracted from the ovary 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.