Figure Legend
Figure 1. Representative specimens of the species used to
design the tetraplex digital PCR assay in this study; A) Burmese python
(Python bivittatus ), B) North African rock python (Python
sebae ), C) rainbow boa (Epicrates cenchria ) and D) boa
constrictor (Boa constrictor ).
Figure 2. Chip images for the multiplex assay when screened
against all four snake species (x-axis) with reporters/probes (y-axis)
showing positive amplification for their corresponding species; red
pixels = positive JUN florescence, yellow pixels = positive ABY
florescence, green pixels = positive VIC florescence, blue pixels =
positive FAM florescence, black pixels = no florescence/negative wells.
Figure 3. Absolute quantification of target DNA for each snake
species when screened with tetraplex assay in mixed samples representing
all possible combinations of multiple species present in a single
sample.
Figure 4. Established populations of Burmese pythons (green),
northern African pythons (blue), boa constrictors (purple), and rainbow
boas (yellow) are shown for Miami-Dade County (blue outline) along with
areas of range overlap among these species. Polygons representing each
constrictor species were created using data available from EDDMapS
(2024) using ArcGIS Pro, Version 3.2 and represent minimum bounding
geometry.
Table 1. Primers and
probes used for generating COI sequences and running qPCR and dPCR
assays for optimization and developing the multiplex assay with
corresponding critical PCR parameters.