The barcode gene sequences should show less intraspecific gene variation than interspecific variation, contain conserved flanking regions and have a short length.
The three most important constraints to explore the suitability of a nucleotide sequence in order to be used as a DNA barcode for taxonomic identifications are:
- Barcode gene sequences show less genetic variation within species (intraspecific variation) than between species (interspecific variation).
- Barcode sequences also show conserved flanking regions in order to be amplified in all species to be identified/categorized.
- Barcode sequences have short lengths due to technical constraints (amplification by PCR is ideal no more than 1000 base pairs).
For example, in animals, an excellent barcode gene sequence used for taxonomic identification is the Cytochrome Oxidase I (COI) gene because this sequence can be used across the evolutionary spectrum.
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