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

  • Pearson G
  • Lago-Leston A
  • Valente M
 et al. 
  • 105

    Readers

    Mendeley users who have this article in their library.
  • 34

    Citations

    Citations of this article.

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 mg
gÀ1 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 À70 C to À80 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.

Author-supplied keywords

  • Brown algae
  • Fucus
  • Lyophilization
  • Plastid gene expression
  • RNA extraction
  • Seagrass
  • rbcl

Get free article suggestions today

Mendeley saves you time finding and organizing research

Sign up here
Already have an account ?Sign in

Find this document

Authors

  • Gareth PearsonCentre of Marine Sciences, University of Algarve

    Follow
  • Asuncion Lago-Leston

  • Marta Valente

  • Ester Serrão

Cite this document

Choose a citation style from the tabs below

Save time finding and organizing research with Mendeley

Sign up for free