Canadian Forest Service Publications

Extraction, detection and persistence of extracellular DNA in forest litter microcosms. 2004. England, L.S.; Vincent, M.; Trevor, J.T.; Holmes, S.B. Molecular and Cellular Probes 18: 313- 319.

Year: 2004

Issued by: Great Lakes Forestry Centre

Catalog ID: 28589

Language: English

CFS Availability: PDF (request by e-mail)

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Abstract

A DNA extraction method was developed that preferentially extracted extracellular DNA rather than intracellular DNA from forest litter. The method purposely avoided the use of harsh chemicals and physical disruption steps used in total DNA extraction to release DNA from cells. The detection limit of PCR, determined by spiking forest litter samples with a dilution series of Choristoneura fumiferana MNPVegt-/lacZ+ genomic DNA, was about 1 ng DNA or 6.85×106 target copies 0.5 g-1 moist forest litter or 0.14 g-1 dry forest litter. In this study, outdoor terrestrial microcosms, each spiked with 49.2 µg of genomic DNA (from the baculovirus CfMNPVegt-/lacZ+), were exposed to summer conditions. A 530 bp DNA fragment from the genome of the baculovirus CfMNPVegt-/lacZ+ was detected in these microcosms for about 3 months. The DNA may have persisted for a longer period but was below the detection limit of the PCR analysis.