title: RNA-seq Chamelea gallina
design: Total RNA was extracted using RNeasy Mini Kit (Qiagen, Hilden, Germany). RNA concentration and quality were assessed by Agilent 2100 Bioanalyzer (Agilent Techologies, Santa Clara, CA, USA.  The cDNA libraries were constructed using a Sure Select Strand-Specific mRNA Library (Agilent Technologies, Santa Clara, CA,USA) according to the manufacturer's protocols. Briefly, before fragmentation, oligo d(T) beads were used to purify poly(A) mRNA from total RNA. First-strand cDNA was synthesized from the fragmented mRNA using random hexamer primers and the cDNA libraries were prepared in accordance with the Illumina protocol. After a purification step the libraries were quantified with a Qubit Fluorometer (Invitrogen, Carlsbad, CA, USA) and pooled together according to their relative concentrations. Its concentration and quality was assesses by Agilent 2100 Bioanalyzer.  The library pool was sequenced on  a HighSeq 4000 (Illumina, Davis, CA, USA) with a single 1x50 bp setup using Version2 chemistry