We aimed to design a versatile PCR primer within the 658 bp COI barcoding region which could be used in combination with a published primer commonly used for DNA barcoding (i.e. LCO1490 or HCO2198
[30 (link)]) to target a short DNA fragment. The Moorea BIOCODE project provided an alignment of 6643 COI sequences belonging to ~3877 marine taxa, mostly coral reef associated species (up to five specimens per morphospecies) spanning 17 animal phyla (sequences available in BOLD, projects MBMIA, MBMIB and MBFA). The information content [entropy h(x)] at each position of the alignment was plotted using BioEdit
[38 ] to locate more conserved regions within the 658 bp COI barcoding fragment (Figure
1 ). A site with limited variation was located between positions 320 and 345 of the 658 bp COI region (Figure
1 ). The forward primer “mlCOIintF” and its reverse complement “mlCOIintR” (Table
1 ) were designed herein and used for further performance testing.
[30 (link)]) to target a short DNA fragment. The Moorea BIOCODE project provided an alignment of 6643 COI sequences belonging to ~3877 marine taxa, mostly coral reef associated species (up to five specimens per morphospecies) spanning 17 animal phyla (sequences available in BOLD, projects MBMIA, MBMIB and MBFA). The information content [entropy h(x)] at each position of the alignment was plotted using BioEdit
[38 ] to locate more conserved regions within the 658 bp COI barcoding fragment (Figure
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