title: RNA-Seq of Mytilus galloprovincialis' digestive gland exposed to Carbamazepine
design: Total RNA was extracted from 5 pools of mussels digestive glands (5 samples each pool) per each experimental group using RNeasy Mini Kit (Qiagen, Hilden, Germany) following manufacturers protocol and with additional DNAse treatment (Qiagen).  RNA purity, concentration and integrity were checked using Qubit Fluorometer (Invitrogen, Carlsbad, CA, USA) and Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA). RNA Integrity Number (RIN) index was calculated for each pool using Agilent 2100 Expert software. Sure Select Strand-Specific mRNA Library (AgilentTechnologies, Santa Clara, CA, USA) was used for the libraries construction according to manufacturer's protocol. Briefly, the first step of RNA library preparation method was the purification of poly(A) RNA from total RNA  using  two serial rounds of binding to oligo(dT) magnetic particles. After  the chemical fragmentation of poly(A) RNA, single-stranding cDNA, the double-stranding cDNA was obtained through the Adenylate cDNA3-ends and the ligate adaptors adding. The final step consisted in the adaptor-ligation cDNA library using the PCR amplification. The libraries were quantified with a Qubit Fluorometer (Invitrogen, Carlsbad, CA, USA) and the quality was assessed by Agilent 2100 Bioanalyzer. The library pools were sequenced on 2 lanes of a HighSeq 4000 (Illumina, Davis, CA, USA) with a single 1*100 bp setup.