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large amounts of PCR-inhibitors. In order to succeed, we first built an ITS2 database 
based on leaf samples and used Sanger barcodes to identify most of the 89 sapwood 
samples to at least family level. However, we were not able to use the same approach 
for the remaining sapwood samples because there were too many problems with 
DNA amplification and sequencing. We therefore switched to NGS barcoding of a 
short trnL gene fragment for 169 sapwood samples. The advantage of this approach 
is that the shorter fragments are more likely to amplify, but this comes at the cost 
of these fragments containing less information. Using this approach, we were able 
to identify 44, and 29 samples with high confidence to the family and genus levels 
respectively. Compared to barcoding of leaf samples, sapwood samples will remain 
very problematic, hence new approaches should continue to be pursued. Particularly 
promising may be anchored hybrid enrichment of chloroplast genes. 
Conclusions 
Being able to identify specimens to species is important for most in-depth study 
of biological systems. However, obtaining these identifications is very challenging 
in tropical environments. Fortunately, a number of new tools make this task less 
daunting. New imaging techniques help with illustrating relevant characters and new 
and cheaper DNA barcodes allow for the generation of databases that can be used by 
many researchers. Overall, making the fauna and flora of Nee Soon freshwater swamp 
forest and Singapore identifiable is achievable. Several hundred, or even thousands of 
species may potentially be revealed from samples that have been collected and stored. 
By focusing on particular taxa belonging to different ecological guilds, it is feasible 
to start understanding species turnover rates across habitats in Singapore and to use 
this information for conserving Singapore’s native fauna and flora. A particularly high 
priority is obtaining plant barcodes for all of Singapore’s vascular plant species. This 
will allow for in-depth studies of species interactions between plants and animals (e.g. 
pollination). 
ACKNOWLEDGEMENTS. This study forms part of the research project “Nee Soon Swamp 
Forest Biodiversity and Hydrology Baseline Studies—Phase 2” funded by the National Parks 
Board (NParks), Singapore. The authors are also grateful to the Public Utilities Board (PUB), 
Singapore for making this study possible. 
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