4.7 Article

Preservation of RNA and DNA from mammal samples under field conditions

期刊

MOLECULAR ECOLOGY RESOURCES
卷 13, 期 4, 页码 663-673

出版社

WILEY
DOI: 10.1111/1755-0998.12108

关键词

degradation; field sampling; NAP buffer; RNAlater; tissue storage

资金

  1. Spanish Ministry of Science and Innovation [CGL-11123, CGL2010-21524, BES-2011-049186]
  2. Spanish Ministry of Education, Culture and Sports [AP2010-5373]

向作者/读者索取更多资源

Ecological and conservation genetics require sampling of organisms in the wild. Appropriate preservation of the collected samples, usually by cryostorage, is key to the quality of the genetic data obtained. Nevertheless, cryopreservation in the field to ensure RNA and DNA stability is not always possible. We compared several nucleic acid preservation solutions appropriate for field sampling and tested them on rat (Rattus rattus) blood, ear and tail tip, liver, brain and muscle. We compared the efficacy of a nucleic acid preservation (NAP) buffer for DNA preservation against 95% ethanol and Longmire buffer, and for RNA preservation against RNAlater (Qiagen) and Longmire buffer, under simulated field conditions. For DNA, the NAP buffer was slightly better than cryopreservation or 95% ethanol, but high molecular weight DNA was preserved in all conditions. The NAP buffer preserved RNA as well as RNAlater. Liver yielded the best RNA and DNA quantity and quality; thus, liver should be the tissue preferentially collected from euthanized animals. We also show that DNA persists in nonpreserved muscle tissue for at least 1week at ambient temperature, although degradation is noticeable in a matter of hours. When cryopreservation is not possible, the NAP buffer is an economical alternative for RNA preservation at ambient temperature for at least 2months and DNA preservation for at least 10months.

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