Deep-Sea Coral Collection Protocols - NOAA's Coral Reef ...

3. Dimethyl sulfoxide (DMSO; 95%)In a population genetic study with a gorgonian in the Caribbean, DMSO worked 90%of the time versus 50% of the time using ethanol. DMSO is non-flammable and nontoxicbut it penetrates the skin, so take precautions against contact with contaminants ortoxics that may use the DMSO medium to penetrate your skin. DMSO is often used inmedicines as a way to slowly release an active component (sticky patches for seasickness, bladder infection treatments). This saturated saline solution is preferred forfixing DNA, but this is a difficult concoction.i. DMSO: 250 ml 0.5M EDTA pH=8.0 100 ml DMSO (final concentration 20%)ii. 105 g NaCl ddH20 for about 400 - 450 ml, Stir a couple of hours, add 10 g ofNACL and stir overnightiii. Next day complete volume to 500 ml and let the excess salt to precipitate.iv. 1.5 ml for 0.5 cm3 of sample, use gloves and prevent contact with skinThere is some question about our ability to get good DNA out of samples that havebeen stored longer than 5 years in DMSO. Some researchers at the prefer DMDMHydrantonin as a non-toxic, non-flammable, long-term solution for tissue preservation.RNA Analysisa. RNA is much less stable than DNA. Various RNA specialty companies market buffers.b. Alhough immediate freezing (but not DMSO) can adequately protect RNA, many researchersuse “RNAlater” (Ambion, Austin Texas, 512-651-0200) to store tissue samples for subsequentlab work.c. RNA will degrade after < 1yr, even if stored at –80. if you want to use RNA down the line,RNA Later is essential..Fig. 26. Primnoid corals in the genus Narella, from Hawaii's deep coral beds.32