title: Ptychodera flava Transcriptome or Gene expression
design: Total RNA was extracted using the Trizol reagent (Invitrogen) from unfertilized eggs and embryos/larvae stages including early blastula, late blastula, early gastrula, late gastrula and tornaria larva. Equal quantities of RNA (20 ug) from eggs and each developmental stage were mixed together, except for the late blastula stage, which was only 10 ug. Subsequently, 50 ug of this pooled total RNA sample was used for the cDNA library construction by Mission Biotech. PolyA-RNA was purified using the Poly(A)Purist MAG Kit (Ambion) and fragmented by RNA fragmentation solution. Double-stranded cDNA was synthesized using the cDNA Synthesis System Kit (Roche) with random primers as described in the protocol provided by the manufacturer. Subsequently, cDNA was purified using the AMPure beads (Agencourt). Sequencing was performed on one pico-titer plate following the protocols for the Genome sequencer GS FLX Titanium System (Roche Diagnostic).