first_imgFifteen species of marine invertebrate commonlyoccurring in the near-shore environment of Rotherabase, Antarctica, were used to test tissue sample storageprotocols with regard to preservation of RNA integrity.After animal collection, the tissues were either immediatelyextracted for RNA or stored at -80C after havingbeen, either directly flash frozen in liquid nitrogen orpreserved in a commercial RNA storage solution, forextraction in the UK. In four cases, direct flash freezingproduced enhanced RNA integrity compared with samplesin the commercial storage solution. A subset of sampleswere further tested for the preferred temperature of storagein the commercial reagent. RNA integrity was well preservedat both ?4 and -20C over periods of 2 months,but degradation was rapid in tissues stored at room temperature.Eight out of the fifteen species only produced asingle ribosomal band on gel electrophoresis. This surveyprovides a guide for tissue transport of Polar cold watermarine invertebrates.Keywords Tissue preservation Tissue transport28 s ribosomal RNA Echinoderms MolluscsIntroductionRNA preservation is sometimes problematic in non-modelspecies but this is particularly the case when dealing withenvironmental species. Logistical issues often surround theability to effectively preserve field-collected samples forRNA analyses. Whilst rapid flash freezing in liquid nitrogengenerally solves this problem, it is not often availablebecause of the remote nature of the work. Even when sucha facility is available on site at a field station, it usuallycannot be transported to the actual, more remote specimencollection site. Also, -80C storage may not be possibleduring transportation from the field site to the mainresearch institute, often thousands of miles away. Antarcticspecimens have the additional issue of operating at temperatures that most species would consider cold and hencecool stow is less effective at reducing tissue degradationthan with, for example, those taken from mammalianspecies. Hence, we decided to carry out a study of effectivestorage protocols for the most common invertebrates foundin the near-shore marine environment in Marguerite Bayclose to Rothera research station, Antarctica.last_img

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