Skip to main content

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

Download Article:
 Download
(PDF 229.2646484375 kb)
 
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.
No References for this article.
No Supplementary Data.
No Article Media
No Metrics

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

Document Type: Research Article

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

Publication date: 2006-02-01

  • Access Key
  • Free content
  • Partial Free content
  • New content
  • Open access content
  • Partial Open access content
  • Subscribed content
  • Partial Subscribed content
  • Free trial content
Cookie Policy
X
Cookie Policy
Ingenta Connect website makes use of cookies so as to keep track of data that you have filled in. I am Happy with this Find out more