title: Recovering 2
design: RNA was extracted from 20-mg Adineta vaga using TRIzol Reagent per the manufacturers protocol, with the addition of linear acrylamide as a co-precipitant.  RNA was quantified with RiboGreen and qualified by 260/230 ratio and Bioanalyzer RNA 6000 Pico Kit.  Libraries were constructed using the KAPA Stranded mRNA-Seq Kit with Illumina TruSeq indexed adapters from 625 ng total RNA per sample.  Magnetic bead binding steps in the library preparation protocol were performed in 1.5-mL tubes.  Final libraries were assessed with PicoGreen and Bioanalyzer High-Sensitivity DNA chip prior to pooling and size selecting 445 bp using a Pippin Prep 1.5% agarose gel cassette.  Final quantification of amplifiable library fragments was done by qPCR.