title: Bcat1 insights into early development of planarians: Spol embryos RNAseq 96h (4 days)
design: At 96 hours post-deposition, egg capsules were placed in a microscope concave glass slide and were treated with 1% diluted commercial NaOCl, for no more than 3 min in order to remove the mucus and parasites. Then the cocoons were moved to an eppendorf tube, washed with RNAlater (Invitrogen) and 50% RNAlater/Trizol. Finally, liquids were removed and 100ul of Trizol reagent (Invitrogen) was added. The whole cocoons were triturated with a tissueLyser (Kontes Microtube Pellet Pestle Rods with Motor), and total mRNA extraction was performed as described in Almuedo-Castillo et al. (2014). Three biological replicates were analyzed per time point. Each biological replicate contained 4 whole cocoons. Library preparation and sequencing were carried out by the Centre Nacional dAnalisi Genomica (CNAG). TruSeq stranded mRNA-seq libraries were sequenced on HiSeq 2000 (Illumina) with a read length of 2x100bp.