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.
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