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Protocol for genotyping cephalopod sex using a skin swab and quantitative PCR
Frederick A Rubino1, Connor J Gibbons2, Thomas J Mungioli2
1Department of Biology, Whitehead Institute, Massachusetts Institute of Technology, Cambridge, MA, USA; The Mortimer B. Zuckerman Mind Brain Behavior Institute, Department of Neuroscience, Columbia University, New York, NY, USA.
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The coleoid cephalopods (octopus, cuttlefish, and squid) are emerging model organisms for neuroscience, development, and evolutionary biology. Determining their sex early in life is critical for population management and controlled experiments. Here, we present a protocol to non-invasively determine the sex of multiple cephalopod species as young as 3 h post-hatching using a skin swab and quantitative PCR (qPCR). We describe steps for designing qPCR primers, swabbing live animals, extracting DNA, running the qPCR, and analyzing the results. For complete details on the use and execution of this protocol, please refer to Rubino et al.1.
