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BioMed Research International
Volume 2014 (2014), Article ID 538549, 9 pages
http://dx.doi.org/10.1155/2014/538549
Research Article

Target Capture and Massive Sequencing of Genes Transcribed in Mytilus galloprovincialis

1Department of Biology, University of Padua, Via U. Bassi 58/b, 32121 Padua, Italy
2Department of Life Sciences, University of Trieste, Via L. Giorgeri, No. 5, 34121 Trieste, Italy

Received 20 February 2014; Revised 29 May 2014; Accepted 7 June 2014; Published 30 June 2014

Academic Editor: Tzu-Ya Weng

Copyright © 2014 Umberto Rosani et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Next generation sequencing (NGS) allows fast and massive production of both genome and transcriptome sequence datasets. As the genome of the Mediterranean mussel Mytilus galloprovincialis is not available at present, we have explored the possibility of reducing the whole genome sequencing efforts by using capture probes coupled with PCR amplification and high-throughput 454-sequencing to enrich selected genomic regions. The enrichment of DNA target sequences was validated by real-time PCR, whereas the efficacy of the applied strategy was evaluated by mapping the 454-output reads against reference transcript data already available for M. galloprovincialis and by measuring coverage, SNPs, number of de novo sequenced introns, and complete gene sequences. Focusing on a target size of nearly 1.5 Mbp, we obtained a target coverage which allowed the identification of more than 250 complete introns, 10,741 SNPs, and also complete gene sequences. This study confirms the transcriptome-based enrichment of gDNA regions as a good strategy to expand knowledge on specific subsets of genes also in nonmodel organisms.