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Whole genome amplification and exome sequencing of archived schistosome miracidia

  • Winka Le Clec'h (a1), Frédéric D. Chevalier (a1), Marina McDew-White (a1), Fiona Allan (a2), Bonnie L. Webster (a2), Anouk N. Gouvras (a2), Safari Kinunghi (a3), Louis-Albert Tchuem Tchuenté (a4) (a5), Amadou Garba (a6), Khalfan A. Mohammed (a7), Shaali M. Ame (a8), Joanne P. Webster (a9), David Rollinson (a2), Aidan M. Emery (a2) and Timothy J. C. Anderson (a1)...
Abstract

Adult schistosomes live in the blood vessels and cannot easily be sampled from humans, so archived miracidia larvae hatched from eggs expelled in feces or urine are commonly used for population genetic studies. Large collections of archived miracidia on FTA cards are now available through the Schistosomiasis Collection at the Natural History Museum (SCAN). Here we describe protocols for whole genome amplification of Schistosoma mansoni and Schistosome haematobium miracidia from these cards, as well as real time PCR quantification of amplified schistosome DNA. We used microgram quantities of DNA obtained for exome capture and sequencing of single miracidia, generating dense polymorphism data across the exome. These methods will facilitate the transition from population genetics, using limited numbers of markers to population genomics using genome-wide marker information, maximising the value of collections such as SCAN.

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Copyright
This is an Open Access article, distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike licence (http://creativecommons.org/licenses/by-nc-sa/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the same Creative Commons licence is included and the original work is properly cited. The written permission of Cambridge University Press must be obtained for commercial re-use.
Corresponding author
Author for correspondence: Winka Le Clec'h, E-mail: winkal@txbiomed.org
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