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Dan Deatherage Kevin Myers September 3,2004

Using Molecular Analysis to Examine the Phylogeny of Symbiotic Water Mites in Two Subgenera: Unionicoloides and Parasitax. Dan Deatherage Kevin Myers September 3,2004. Background. Symbiotic water mites Mussels provide hosts to mites Size requires molecular techniques

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Dan Deatherage Kevin Myers September 3,2004

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  1. Using Molecular Analysis to Examine the Phylogeny of Symbiotic Water Mites in Two Subgenera:Unionicoloides and Parasitax Dan Deatherage Kevin Myers September 3,2004

  2. Background • Symbiotic water mites • Mussels provide hosts to mites • Size requires molecular techniques • Molecular phylogeny Vs. Classical Phylogeny • Cytocrome Oxidase I (COI) gene Vs. Internal Transcriber Sequence (ITS)

  3. Why These Mites? • Readily found and collected • Generally many individuals in each population • Mites similar to these have been heavily studied • The COI sequences for these mites are not known • This field of study is open to new research without fear of being “scooped” • The dissemination of species was begun by Dr. Edwards

  4. Unionicola sp. • All species of Unionicola are morphologically very similar Utterbackita imbecillisAnadonta suborbiculata

  5. Proposed Research • Project begun through the UExplore Undergraduate Research Program summer 2004. • Mites chosen because their close classical phylogeny classification. • Results during the summer have shown that the methods we will use are effective.

  6. Species of Mites to Study Include: • From the subgenus Parisitax: • Unionicola formosa from Pyganadon cataracta • U. formosa from P. grandis • From the subgenus Unionicoloides: • Unionicola kavanaughi • U. gailae • U. hoesei • U. lasallei • U. amandita

  7. For Each Mite

  8. Primer Design with NetPrimer

  9. For Each Mite

  10. Ladder |–––––1-4––––––||–––––5-8––––––| |–-––––9-12–––-––| Gel electrophoresis of U. foili from U. imbecillis (1-8) and U. formosa from A. suborbiculata (7-12). There are bright bands for each mite, supporting the near correct sequence of our primers.

  11. For Each Mite

  12. CodonCode Aligner

  13. For Each Mite

  14. ClustalW

  15. Molecular Phylogenic Tree

  16. Timeline • Sept 15 Preliminary extraction underway. All materials gathered. • Sept 30 All DNA extracted from initial mites and 2 species sent for sequencing. • Oct 15 2 more species sequenced, initial 2 species sequence analysis began. • Oct 30 Final 2 sequences returned, analysis continuing. • Nov 15 All sequence analysis complete, manuscript begun. • Nov 30 All lab work done, and manuscript in final stages. • Dec 15 Manuscript submitted for class and for publication.

  17. Budget ItemCost • DNA Sequencing $300 (16 sequences) • Qiagen DNeasy Tissue Kit $156 • Promega PCR Master Mix $67 (100 reactions) Total: $523

  18. Grade Agreement

  19. References • Boore, J. L. 1999. Animal Mitochondrial Genomes. Nucleic Acids Research, 27(8):1767-1780. • Edwards, D. D. and Ernsting, B. R. 2004. UExplore Undergraduate Research Proposal. • Myers, Kevin, Eyler, Andrea, and Rasure, Mark. 2004. Unpublished Data. • Navajas, M. and P. Boursot. 2003. Nuclear ribosomal DNA monoplyly versus mitochondrial DNA polyphyly in two closely related mite species: the influence of life history and molecular drive. Proceedings of the Royal Society of London B Biological Sciences, 270 Suppl 1:S124-127.

  20. Questions? Ideas?

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