title: Preservation of RNA and DNA from mammal samples under field conditions.
abstract: 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 1 week 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 2 months and DNA preservation for at least 10 months.
authors.affiliation: Conservation and Evolutionary Genetics Group, Estación Biológica de Doñana (EBD-CSIC), Seville, Spain. miguel.camacho@ebd.csic.es
authors.forename: Miguel
authors.initials: M
authors.lastname: Camacho-Sanchez
authors.forename: Pablo
authors.initials: P
authors.lastname: Burraco
authors.forename: Ivan
authors.initials: I
authors.lastname: Gomez-Mestre
authors.forename: Jennifer A
authors.initials: JA
authors.lastname: Leonard
mesh_terms: D000825:Animal Structures / Q000737:chemistry; D000818:Animals; D002021:Buffers; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification*; D005615:Freezing; D011309:Preservation, Biological / Q000379:methods*; D012313:RNA / Q000235:genetics* / Q000302:isolation & purification*; D051381:Rats
chemical_list: D002021:Buffers; D012313:RNA; D004247:DNA