title: A novel method for the transport and analysis of genetic material from polyps and zooxanthellae of scleractinian corals.
abstract: We have developed a new simple method for transport, storage, and analysis of genetic material from the corals Agaricia agaricites, Dendrogyra cylindrica, Eusmilia ancora, Meandrina meandrites, Montastrea annularis, Porites astreoides, Porites furcata, Porites porites, and Siderastrea siderea at room temperature. All species yielded sufficient DNA from a single FTA card (19 microg-43 ng) for subsequent PCR amplification of both coral and zooxanthellar DNA. The D1 and D2 variable region of the large subunit rRNA gene (LSUrDNA) was amplified from the DNA of P. furcata and S. siderea by PCR. Electrophoresis yielded two major DNA bands: an 800-base pair (bp) DNA, which represented the coral ribosomal RNA (rRNA) gene, and a 600-bp DNA, which represented the zooxanthellar srRNA gene. Extraction of DNA from the bands yielded between 290 microg total DNA (S. siderea coral DNA) and 9 microg total DNA (P. furcata zooxanthellar DNA). The ability to transport and store genetic material from scleractinian corals without resort to laboratory facilities in the field allows for the molecular study of a far wider range and variety of coral sites than have been studied to date.
authors.affiliation: School of Animal and Microbial Sciences, The University of Reading, PO Box 228, Whiteknights, Berkshire RG6 6AJ, Reading, UK. m.j.c.crabbe@rdg.ac.uk
authors.forename: M James C
authors.initials: MJ
authors.lastname: Crabbe
mesh_terms: D000818:Animals; D037421:Anthozoa / Q000737:chemistry* / Q000166:cytology / Q000235:genetics*; D004247:DNA / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*; D019544:Equipment Failure Analysis; D020869:Gene Expression Profiling / Q000295:instrumentation* / Q000379:methods*; D011786:Quality Control; D013048:Specimen Handling / Q000295:instrumentation* / Q000379:methods*
chemical_list: D004247:DNA