liver_red-spotted_newt; Notophthalmus viridescens; RNA-Seq
RNA-Seq library was prepared using TruSeq RNA Sample Prep Kits v2 (Illumina) according to manufacturers recommendations combined with the dUTP method previously described(25). Additional modifications are: First, we started with 1-5 µg of total RNA. RNA was fragmented for 1 min at 94 oC. After the first strand synthesis, the sample was purified using QIAquick PCR Purification kit (Qiagen) to remove all dNTPS and eluted into 25 µl of RNAse/DNAse free water. Different RNA Adapter Indexes were used to make indexed library. The library was amplified for 12 cycles. The resulting library was size-selected (500 ± 25 bp) using LabChipXT (Caliper) according to manufacturers recommendations.  Finally, resulting libraries were paired-end (2x100 bp) sequenced on an Illumina Hiseq 2000.