Venom research has attracted an increasing interest in disparate fields, from drug
development and pharmacology, to evolutionary biology and ecology, and rational antivenom
production. Advances in “-omics” technologies have allowed the characterization of an increasing
number of animal venoms, but the methodology currently available is suboptimal for large-scale
comparisons of venom profiles. Here, we describe a fast, reproducible and semi-automated protocol
for investigating snake venom variability, especially at the intraspecific level, using the Agilent
Bioanalyzer on-chip technology. Our protocol generated a phenotype matrix which can be used
for robust statistical analysis and correlations of venom variation with ecological correlates, or
other extrinsic factors. We also demonstrate the ease and utility of combining on-chip technology
with previously fractionated venoms for detection of specific individual toxin proteins. Our study
describes a novel strategy for rapid venom discrimination and analysis of compositional variation at
multiple taxonomic levels, allowing researchers to tackle evolutionary questions and unveiling the
drivers of the incredible biodiversity of venoms.