. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. Broken Spur TAG A~ — Snake Pit /_____— Logatchev - ,60°w o° Figure 1. Geographic locations of Bathymodiolus collection sites along the Mid-Atlantic Ridge. Major fracture zones () are shown. Materials and Methods Specimens Using the deep submergence vehicle Alvin 's mechanical claw, samples were collected along the MAR (Fig. 1. Table I) in July 1997. Once aboard the support vessel Atlantis, we placed the specimens in 4°C filtered seawater. Mytilids were then either dissected or frozen whole. For dissected specimens
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. Broken Spur TAG A~ — Snake Pit /_____— Logatchev - ,60°w o° Figure 1. Geographic locations of Bathymodiolus collection sites along the Mid-Atlantic Ridge. Major fracture zones () are shown. Materials and Methods Specimens Using the deep submergence vehicle Alvin 's mechanical claw, samples were collected along the MAR (Fig. 1. Table I) in July 1997. Once aboard the support vessel Atlantis, we placed the specimens in 4°C filtered seawater. Mytilids were then either dissected or frozen whole. For dissected specimens, gill, mantle, adductor muscle, and tissue mass were frozen at — 70°C in labeled bags; shells were then measured and labeled. All samples were brought back to Rutgers University on dry ice and stored at — 80°C until they were prepared for genetic analysis. DNA extraction, polymerase chain reaction, and sequencing Whole cellular DNA was isolated by digesting g of frozen tissue following a hexadecyl-trimethyl-ammo- nium bromide (CTAB) protocol (Doyle and Dickson, 1987), followed by a phenol extraction and ethanol precipitation (Sambrook et ai. 1989). Purified nucleic acids were rehy- drated in IX TE buffer (10 mM Tris-HCl. pH : 1 mM EDTA) to a final concentration of 100-500 ng/jul and stored for short periods at 4°C and for longer periods at — 20°C. An approximately 710-bp region of the mitochondria was amplified using primers designed to amplify ND4-L in fish. Arg BL (5'-CAA gAC CCT TgA TTT Cgg CTC A-3') is in the tRNA arginine (Bielawski and Gold, 1996) and NAP 2H (5'-Tgg AgC TTC TAC gig A/ggC TTT-3') is within ND4 itself (Arevalo et al., 1994). Comparison of the result- ing sequences via BLASTN (Altschul et ai, 1990) and BLASTP (Altschul et al., 1997) searches of GenBank se- quences indicated that these primers amplified tRNA-Met, tRNA-Val, and the first third of ND4 in Bathymodiolus. The amplification reaction contained 100-500 ng tem- pl
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Keywords: ., bookauthorlilliefrankrat, booksubjectbiology, booksubjectzoology
