RNAseq of Philodryadini venom glands
Initially, the total RNA was extracted with Trizol and quantified by Quant-It Ribogreen RNA and Reagent Kit (Invitrogen, Life Technologies Corp., Waltham, MA, USA). Then, the quality control of the extracted RNA integrity was performed in an Agilent 2100 Bioana-lyzer (Santa Clara, California, USA) using an Agilent RNA 6000 Nano kit. Libraries were prepared for each sample. One g of total RNA was used with the Illumina Truseq Stranded RNA HT (Hayward, CA, USA) kit consisting of cDNA Truseq Stranded RNA HT/PCR synthesis, Truseq Stranded RNA HT/Adapter Card and Truseq Stranded HT mRNA. The libraries were validated considering the distribution of the fragments' size and their quantification. Then, the quantification of each library was performed by Re-al-Time PCR using the KAPA SYBR FAST Universal qPCR kit (Merck, Darmstadt, Hesse, Germany), according to the manufacturer's protocol, using the StepOnePlus  Real-Time PCR System (Waltham, MA, USA). Aliquots of each cDNA library were diluted to a con-centration of 2 nM. Then, a pool of all samples with 5 L from each library was prepared, and the pool concentration was again determined by real-time PCR. cDNA libraries were sequenced in the Illumina HiSeq 1500 System (Hayward, CA, USA) in a rapid paired end flowcell in 300 paired end-end strategy cycles of 2  150 bp.