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Fast and easy method for total DNA extraction and gene amplification from larvae,spat and adult mussels Mytilus trossulus from the Baltic Sea
Authors:Rafał Lasota  Joanna Piłczyńska  Suzanne T. Williams  Maciej Wołowicz
Affiliation:1. Laboratory of Estuarine Ecology, Institute of Oceanography, University of Gdańsk, Al. M. Pi?sudskiego 46, 81-372, Gdynia, Poland
3. Department of Life Sciences, Natural History Museum, Cromwell Rd, London, SW7 5BD, UK
Abstract:Three nuclear DNA markers that diagnostically differentiate mussels within the Mytilus edulis complex (M. edulis, M. trossulus and M. galloprovincialis) are commonly used in taxonomic investigations: Glu5’, ITS and EFbis. As a rule, DNA extraction is performed before amplification. It is a time consuming process in the case of traditional methods based on chloroform and phenol extraction or relatively expensive using kits with ready spin columns. Moreover, DNA isolation from larvae is problematic, because of the small amount of tissue available. In this report we describe a simple, fast and inexpensive method of DNA extraction and gene amplification from larvae, spat and adults of the Baltic mussel Mytilus trossulus. The extraction method is adapted from that of Wang et al. (2006) and is based on digestion of tissue or whole animals in STE solution and direct gene amplification. On the basis of the results of routine analyses of mussels carried out in our laboratory we have concluded that the method we propose gives results that are consistent with standard methods, without requiring expensive reagents/equipment and is time saving.
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