Amphiura arm 95% DI tissue
RNA was extracted from 95% DI arms with RiboPure(Ambion). Total RNA (800 ng) was used for cDNA synthesis using the SMART approach (Mint-Universal cDNA synthesis kit, Evrogen). An aliquot of 100 ng of cDNA was subsequently normalized using duplex-specific nuclease and re-amplified afterwards according to the manufacturer's instructions (Trimmer kit, Evrogen). Libraries for 454 sequencing were prepared from cDNA with the ‘GS FLX Titanium General Library Preparation Kit’. The 454 libraries so obtained were immobilized on beads and clonally amplified using the ‘GS FLX Titanium LV emPCR Kit (Lib-L)’. The libraries were then sequenced using the ‘GS FLX Titanium Sequencing Kit XLR70’ and ‘GS FLX Titanium PicoTiterPlate Kit’.