Fast and easy method for total DNA extraction and gene amplification from larvae, spat and adult mussels Mytilus trossulus from the Baltic Sea

• Autorzy: Lasota, R. , Piłczyńska, J. , Williams, S. T. , Wołowicz, M.
• Języki publikacji: EN

Abstrakty

EN

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.

Słowa kluczowe

EN

Mytilus trossulus; DNA extraction method; molecular markers; PCR

• Czasopismo: Oceanological and Hydrobiological Studies
• Rocznik: 2013
• Tom: Vol. 42, No. 4
• Strony: 486--489
• Opis fizyczny: Bibliogr. 7 poz., rys.

Twórcy

autor

Lasota, R.

• Laboratory of Estuarine Ecology, Institute of Oceanography, University of Gdańsk, Al. M. Piłsudskiego 46, 81-372, Gdynia, Poland,

autor

Piłczyńska, J.

• Laboratory of Estuarine Ecology, Institute of Oceanography, University of Gdańsk, Al. M. Piłsudskiego 46, 81-372, Gdynia, Poland

autor

Williams, S. T.

• Department of Life Sciences, Natural History Museum, Cromwell Rd, London, SW7 5BD, UK

autor

Wołowicz, M.

• Laboratory of Estuarine Ecology, Institute of Oceanography, University of Gdańsk, Al. M. Piłsudskiego 46, 81-372, Gdynia, Poland

Bibliografia

• 1. Aucoin F., Doiron S., Nadeau M., (2004). Guide to sampling and identifying larvae of species of maricultural interest. Guide publication G005. Gaspe (Canada): MAPAQ. 73 pp. ISBN: 1-55137-600-8
• 2. Beaumont, A. R., Hawkins M. P., Doig F. L., Davies I. M., Snow M., (2008). Three species of Mytilus and their hybrids identified in a Scottish Loch: natives, relicts and invaders? J. Exp. Mar. Biol. Ecol. 367(2): 100–110. DOI:10.1016/j.jembe.2008.08.021
• 3. Heath D. D., Rawson P. D., Hilbish T. J., (1995). PCR-based nuclear markers identify alien blue mussel (Mytilus spp.) genotypes on the west coast of Canada. Can. J. Fish. Aquat. Sci. 52:2621–2627. DOI:10.1139/f95-851
• 4. Inoue, K., Waite H., Matsuoka M., Odo S., Harayama S., (1995). Interspecific variations in adhesive protein sequences of Mytilus edulis, M. galloprovincialis, and M. trossulus. Biol. Bull. 189:370–375. DOI: 10.2307/1542155
• 5. Kijewski T., Zbawicka M., Väinölä R., Wenne R., (2006). Introgression and mitochondrial DNA heteroplasmy in the Baltic populations of mussels Mytilus trossulus and M. edulis. Mar. Biol. 149:1371–85. DOI:10.1007/s00227-006-0316-2
• 6. Wang S., Bao Z., Zhang L., Li N., Zhan A., Guo W., Wang X., Hu J., (2006). A new strategy for species identification of planktonic larvae: PCR-RFLP analysis of the internal transcribed spacer region of ribosomal DNA detected by agarose gel electrophoresis or DHPLC. J. Plankton Res. 28(4): 375–384. DOI: 10.1093/plankt/fbi122
• 7. Williams S.T., Reid D.G., Littlewood, D.T.J., (2003). A molecular phylogeny of the Littorininae (Gastropoda: Littorinidae): unequal evolutionary rates, morphological parallelism and biogeography of the Southern Ocean. Mol. Phylogenet. Evol. 28: 60–86. DOI: 10.1016/S1055-7903(03)00038-1