An inexpensive and rapid RNA extraction protocol for brown algae and seagrasses is presented, based on homogenization in a simple CTAB buffer and selective precipitation of RNA with lithium chloride. The protocol avoids the use of toxic chaotropic agents and phenol; high concentrations of dithiothreitol are used to inhibit RNase activity and to prevent oxidative cross-linking of nucleic acids by phenolics. A relatively high throughput of about 100 samples in 24 h can be achieved for, for example, Northern analysis. Yields of 50-200 μg g-1 fresh weight are comparable with those obtained for higher plants using commercially available kits. Tests of the extraction procedure demonstrate that high quality, intact RNA can be obtained from a variety of lyophilized brown algal tissues, even after prolonged storage at room temperature. Lyophilization is therefore suggested as an alternative to freezing tissue at -70°C to -80°C. The RNA obtained was used directly in several downstream applications to detect Fucus plastid-encoded transcripts by RNA-labelling, RT-PCR and Northern analysis. © 2006 British Phycological Society.
CITATION STYLE
Pearson, G., Lago-Leston, A., Valente, M., & Serrão, E. (2006). Simple and rapid RNA extraction from freeze-dried tissue of brown algae and seagrasses. European Journal of Phycology, 41(1), 97–104. https://doi.org/10.1080/09670260500505011
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