GSM2187941: e4_1; Schmidtea mediterranea; RNA-Seq
Live embryos were dissected out of egg capsules in 1x Holfreter’s buffer (3.5 g/L NaCl; 0.2 g/L NaHCO3; 0.05 g/L KCl; 0.2 g/L MgSO4; 0.1 g/L CaCl2; 1.0 g/L dextrose, pH 7.0-7.5) for Stage (S) S2-S7 egg capsules, or 1x Montjuic water for S8 hatchlings.  Yolk samples were obtained from 8 d egg capsules that contained neither spherical nor elongating embryos.  Single embryos were  transferred into microfuge tubes containing 200 µl TRIzol reagent (Thermo Fisher, item #15596-018), homogenized by pipetting, and stored at -80˚C. Single animal samples of intact C4 adults and virgin, sexually mature adults were homogenized in 1.0 mL TRIzol using an IKA Ultra Turrax T 25 Basic tissue disruptor prior to storage at -80˚C.  Total RNA extraction was performed in 1.0 mL TRIzol per sample according to the manufacturer’s protocol, following the recommendations for working with small amounts of tissue. PolyA-selected, single stranded RNA-Sequencing libraries were prepared for four biological replicates per stage using the Illumina TruSeq RNA Sample V2 kit (item # RS-122-2001 and RS-122-2002), starting with 500 ng total RNA per sample (C4, virgin sexual adult (SX), Y, S4, S5, S6, S7, S8), or 100 ng total RNA per sample (S2, S3).
GEO Accession
GSM2187941