Journal of Applied Phycology

, Volume 17, Issue 3, pp 223–229 | Cite as

Development of new procedures for the isolation of phytoplankton DNA from fixed samples

  • Elena Bertozzini
  • Antonella Penna
  • Elisa Pierboni
  • Ian Bruce
  • Mauro Magnani
Article

Abstract

Phytoplankton samples collected for routine monitoring programmes have traditionally been preserved with fixatives before subsequent analytical procedures such as microscope-based identification, or simply to permit transport between laboratories. In recent years, to simplify identification and enumeration, the use of DNA or RNA probes coupled with the PCR assay has progressed and now represents a routine procedure for screening cultured and field samples. However, the phytoplankton cells have often still to be treated as fixed samples.

The extraction of genomic DNA from fixed cultures of Alexandrium minutum cultures was compared using two new methods based on Magnetisable Solid Phase Support (MSPS) techniques with that using three commercial kits. Genomic DNA recovery and PCR amplification were observed and the results obtained from culture samples were validated using field samples. Among the DNA extraction techniques considered, the MSPS methods provided the best results.

Key Words

DNA extraction fixatives harmful algal blooms (HABs) magnetisable solid phase support (MSPS) PCR 

Abbreviations

DEAE

Diethylaminoethyl group

HAB

Harmful Algal Bloom

ITS

Internal transcribed spacer

MSPS

Magnetisable Solid Phase Support

PEG

polyethylene glycol

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Copyright information

© Springer Science + Business Media, Inc. 2005

Authors and Affiliations

  • Elena Bertozzini
    • 1
  • Antonella Penna
    • 2
  • Elisa Pierboni
    • 3
  • Ian Bruce
    • 4
  • Mauro Magnani
    • 5
  1. 1.Centre of BiotechnologyUniversity of UrbinoFanoItaly
  2. 2.Centro Biologia AmbientaleUniversity of UrbinoPesaroItaly
  3. 3.Istituto Zooprofilattico Sperimentale Umbria e MarchePerugiaItaly
  4. 4.School of Chemical and Life SciencesUniversity of GreenwichKentU.K.
  5. 5.Institute of Biological Chemistry “G. Fornaini”University of UrbinoUrbinoItaly

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