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. 2012 Jun 29;13:287. doi: 10.1186/1471-2164-13-287

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Copyright ©2012 Nyberg et al.; licensee BioMed Central Ltd.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Workflow of cDNA library construction. A mixed-stage regeneration/fission cDNA library was generated from ~4,500 P. leidyi worms. Anteriorly and posteriorly regenerating worms were collected from 0 to 3.5 days after amputation (dotted lines mark amputation planes; gray terminal masses represent regeneration blastemas) and actively fissioning worms were also collected (gray shading marks intercalated head and tail tissue that forms during fission). Following RNA extraction and cDNA synthesis, a portion of the pooled cDNA was normalized. The final library sent for 454 sequencing consisted of 2/3 normalized and 1/3 non-normalized cDNA.