title: Rehydration conditions for isolation of high quality RNA from the lichen Lobaria pulmonaria.
abstract: BACKGROUND
  The poikilohydric nature of lichens enables them to survive repeated episodes of desiccation by utilizing water when it becomes available. During rehydration, RNA-degrading endonucleases may be released, reducing RNA quantity and quality. Re-generation of a steady-state condition where RNA quantity and quality no longer fluctuate establishes a framework for development of new hypotheses for future investigations. Using Lobaria pulmonaria as a model species, the objective of this study was to compare the effect of different rehydration conditions on the quantity and quality of RNA from the rehydrated thallus.
  
  
  RESULTS
  Spectrophotometric measurements of total RNA and cDNA were performed for samples prepared from dry lichen or lichen after rehydration (0.5 h, 1 h, 2 h, 4 h or 24 h), with limited light and dark conditions, and at three temperatures (15°C, 20°C or 32°C) for some of these conditions. The results showed that rehydration of the thallus for 4 h at 20°C in light yielded the highest concentration and quality of RNA. A higher RNA concentration was obtained in light than in dark conditions, but the RNA quality was unaffected.
  
  
  CONCLUSIONS
  This study suggests that allowance of 4 h for thallus rehydration should be adequate to ensure complete recovery of transcription. After 4 h at 20°C further studies can be carried out on the RNA in this model species.
authors.forename: Jennifer A
authors.initials: JA
authors.lastname: Doering
authors.forename: Vivian P W
authors.initials: VP
authors.lastname: Miao
authors.affiliation: Department of Biological Sciences, University of Manitoba, 50 Sifton Road, Winnipeg, MB R3T 2 N2, Canada. Michele.Piercey-Normore@umanitoba.ca.
authors.forename: Michele D
authors.initials: MD
authors.lastname: Piercey-Normore
mesh_terms: D003890:Desiccation; D008011:Lichens / Q000737:chemistry* / Q000378:metabolism; D008027:Light; D020871:RNA Stability / Q000528:radiation effects; D012331:RNA, Fungal / Q000302:isolation & purification*; D013048:Specimen Handling; D013696:Temperature; D013997:Time Factors; D014867:Water / Q000737:chemistry*
chemical_list: D012331:RNA, Fungal; D014867:Water