Rehydration conditions for isolation of high quality RNA from the lichen Lobaria pulmonaria

BMC Research Notes

Michele D Piercey-Normore²  Vivian PW Miao¹  Jennifer A Doering² 

[1] Department of Microbiology and Immunology, University of British Columbia, 2350 Health Sciences Mall, Vancouver, BC V6T 1Z3, Canada;Department of Biological Sciences, University of Manitoba, 50 Sifton Road, Winnipeg, MB R3T 2 N2, Canada

关键词: Total RNA;    RT-PCR;    RNA quantity;    RNA quality;    Lobaria pulmonaria;    Lichen;    cDNA;   

Others  :  1131971
DOI  :  10.1186/1756-0500-7-442

 received in 2013-12-11, accepted in 2014-07-08,  发布年份 2014

【 摘 要 】

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.

Findings

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.

【 授权许可】

   
2014 Doering et al.; licensee BioMed Central Ltd.

【 图 表 】

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