Abstract
Molecular genetic markers have numerous potential applications in environmental forensics
if DNA can be isolated from ‘difficult’ non-human biological material such as hairs,
feathers, or wood. The identification of the origin of wood is particularly important
in order to identify illegally harvested and traded timber and wood products. We describe
success rates of DNA isolation from wood based on a simple, previously published extraction
protocol. The protocol was used to isolate DNA from a total of 406 wood samples, mainly
of the important tropical tree family Dipterocarpaceae. The reliability of the extraction
method was confirmed by comparing fragment sizes and sequences after isolation of
DNA from leaves and wood of the same trees. We observed the success of amplification
of chloroplast DNA (cpDNA) fragments of different lengths by means of PCR, investigated
key factors influencing PCR, and conducted inhibitor tests for a subset of the samples.
The average rate of successful PCR amplification was 75.7%. Main factors influencing
the success of PCR amplification were the size of the amplified fragment and the processing
status of the wood. Short fragments and unprocessed wood resulted in higher success
rates. The success rate was also dependent on the age (storage duration) of the wood
probe and on the investigated species. Amplification success was higher if DNA was
isolated from outer sapwood (without cambium) in comparison to DNA isolated from the
transition zone between sapwood and heartwood and the inner heartwood. However, inhibitor
tests also indicated more PCR inhibitory substances in the outer sapwood in comparison
to transition wood and heartwood. The addition of polyvinylpyrolidone (PVP) to the
lysis buffer proved to be highly efficient to improve the amplification success if
inhibitory substances were present.
Keywords
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Article info
Publication history
Published online: February 05, 2009
Accepted:
January 6,
2009
Received in revised form:
December 17,
2008
Received:
September 27,
2008
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Copyright
© 2009 Elsevier Ireland Ltd. Published by Elsevier Inc. All rights reserved.
