Transcriptome from Phoronis australis
Tissue  were preserved in at least 10 volumes of RNAlater® soon after the animals were collected; if sent to the laboratory alive, animals were flash-frozen in liquid nitrogen. All samples were stored at -80 ºC until RNA was extracted. Total RNA was extracted using Tri-Reagent® (Ambion), following the manufacturer’s protocol. Subsequent mRNA purification was performed with the Dynabeads® mRNA Purification Kit for mRNA (Invitrogen). Purification from Total RNA preps followed the manufacturer’s instructions. Further details of the RNA extraction and purification protocols can be found in  Riesgo, Andrade, et al. (2012) .   Quantity and quality (purity and integrity) of mRNA were assessed by two different methods. Quantity of mRNA was measured with the fluorometric quantitation performed by the QubiT® Fluorometer (Invitrogen, California, USA). Also, capillary electrophoresis in an RNA Pico 6000 chip was done using an Agilent Bioanalyzer 2100 System with the “mRNA pico Series II” assay (Agilent Technologies, California, USA). Integrity of mRNA was estimated by the electropherogram profile and lack of rRNA contamination (based on rRNA peaks for 18S and 28S rRNA given by the Bioanalyzer software).