Simple and rapid RNA extraction from freeze-dried tissue of brown algae and seagrasses

Authors: Pearson, Gareth; Lago-Leston, Asuncion; Valente, Marta; Serrão, Ester

Source: European Journal of Phycology, Volume 41, Number 1, February 2006 , pp. 97-104(8)

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Abstract:

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.

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Keywords: brown algae; Fucus; lyophilization; plastid gene expression; rbcL; RNA extraction; seagrass

Document Type: Research article

DOI: http://dx.doi.org/10.1080/09670260500505011

Affiliations: 1: CCMAR, CIMAR/Laboratório Associado, F.C.M.A., Universidade do Algarve, Gambelas, 8005–139 Faro, Portugal

Publication date: 2006-02-01