Using Next Generation Amplicon Sequencing to undertake a high resolution diet study in true sea

snakes

Blanche D’Anastasi – PhD Candidate1,2,3

Dr Vimoksalehi Lukoschek – Research Fellow1

Dr Lynne van Herwerden – Senior Lecturer2

Dr. Colin Simpfendorfer – Professor and Centre Director3,4

Dr. Jean-Paul Hobbs – Post Doctoral Fellow5

1Australian Research Council Centre of Excellence for Coral Reef Studies, JCU2School of Marine and Tropical Biology, James Cook University3School of Earth and Environmental Sciences, JCU4Centre for Sustainable Tropical Fisheries and Aquaculture, James Cook University 5Oceans Institute, University of Western Australia

Simon Allen

1. Extract DNA from intestinal content of at least 20 snakes

per species

2. Use PCR to amplify genes of prey items and tag gut contents of 25 snakes

3. Pool equal amount of amplicons from 25 snakesN

GA

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How

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4. Send for Next Generation Sequencing on a Roche GS

Junior 454 platform

NG

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5. A library of 7000 sequence reads per snake

6. Use Bioinformatics platforms to match each read to a data bases

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AS –

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s7. No match – geographic

information or assign higher taxonomic status

8. Calculate % contribution of each prey item

Murray et al., 2011