DNA barcoding for Nee Soon flora and fauna 
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Image plates were exported in a format that allows for online magnification. This 
allows the users on internet browsers to view both an overall view of the plate but also 
zoom in to high-magnification images of structures that are critical for identification: it 
divides an image into a series of smaller-sized picture tiles at different resolutions and 
sizes that are presented onto a fixed frame. Because the viewer frame requires only few 
picture tiles to be loaded at any time, viewing is fast and smooth. 
Online image database. All images are displayed on an online image database 
(https://singapore.biodiversity.online/). The website also displays other species from 
Singapore. The webpage has a collapsible taxon-based navigation panel on the left, 
while the main field displays all imaged specimens (in thumbnails). There are also 
filter options. The image database features a three-tiered design; users can select a 
taxon group on the left navigation panel, which will show all the available species on 
the right panel, segregated by another two taxon levels (usually, order, followed by 
family or genus). Clicking on a species thumbnail will direct users to the species page, 
which displays the image plate for the specimen, as well as other basic information 
such as species name, common name, taxonomic information, image information 
as well as other additional species information and links to other websites for more 
information on the species (where available). Users can then navigate to other taxon 
groups using navigation panels. 
Results 
Faunal barcoding 
Using a combination of traditional Sanger sequencing and high throughput Next 
Generation Sequencing (NGS barcoding, we generated 2904 animal barcodes 
(predominantly COI, with some COII for Odonata) (Table 1). Overall, we have a total 
of 2904 barcodes from the faunal specimens collected from Nee Soon freshwater 
swamp forest. The majority of barcodes are for insects: Diptera (1399), ants (652) 
and Odonata (347). Insects collected from Nee Soon’s waterways are represented by 
128 barcodes. Fishes represent the next largest contribution (201) of barcodes for non¬ 
insect fauna. However, numerous additional barcodes continue to be generated for the 
material that was collected. All barcodes were checked against Genbank in order to 
rule out contamination. 
DNA barcode database for plants 
A total of 1189 barcodes were generated for 503 species representing 98 plant families, 
using a combination of traditional Sanger sequencing and high throughput Next 
Generation Sequencing. We also generated genome data for 240 species from which 
170 chloroplast genomes could be assembled via genome skimming. Coverage of 
chloroplast genomes depended on both sequencing depth and amount of chloroplast 
