Demonstration
of extraction and PCR amplification of DNA from phytoplankton of lakes with
high humic acid content
M. LeBreton, P. Morton, K. Larade, B. Harland, T.
Clair, D. Campbell-2000
Hydrobiologia, 438 (1/3):91-97
Abstract:
Lakes with high levels of dissolved organic matter are common in northern
temperate regions. These habitats are sensitive to environmental and climate
change, but the molecular ecology and eco-physiology of their phytoplankton
have been under studied, in part because the DOM interferes with molecular
and biochemical analyses of these populations. We developed a procedure
which uses polyvinylpyrrolidone to adsorb interfering material during the
extraction of nucleic acids from these populations. This procedure does not
depend on enzymatic lysis and yields DNA of sufficient quality to serve as a
template for PCR amplification of specific genetic sequences. In particular,
we detected fragments of the conserved rbcL gene encoding the large subunit
of the Rubisco enzyme from the phytoplankton of lakes with high DOM. Our
extraction procedure may be useful to other workers studying similar
populations.
(Department of Biology, Mount Allison University,
Sackville, NB, E4L 1G7, Canada, E-mail of D. Campbell: dcampbell@mta.ca)
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