TY - JOUR
T1 - The impact of endogenous content, replicates and pooling on genome capture from faecal samples
AU - Hernandez-Rodriguez, Jessica
AU - Arandjelovic, Mimi
AU - Lester, Jack
AU - de Filippo, Cesare
AU - Weihmann, Antje
AU - Meyer, Matthias
AU - Angedakin, Samuel
AU - Casals, Ferran
AU - Navarro, Arcadi
AU - Vigilant, Linda
AU - Kühl, Hjalmar S.
AU - Langergraber, Kevin
AU - Boesch, Christophe
AU - Hughes, David
AU - Marques-Bonet, Tomas
N1 - Publisher Copyright:
© 2017 The Authors. Molecular Ecology Resources Published by John Wiley & Sons Ltd.
PY - 2018/3
Y1 - 2018/3
N2 - Target-capture approach has improved over the past years, proving to be very efficient tool for selectively sequencing genetic regions of interest. These methods have also allowed the use of noninvasive samples such as faeces (characterized by their low quantity and quality of endogenous DNA) to be used in conservation genomic, evolution and population genetic studies. Here we aim to test different protocols and strategies for exome capture using the Roche SeqCap EZ Developer kit (57.5 Mb). First, we captured a complex pool of DNA libraries. Second, we assessed the influence of using more than one faecal sample, extract and/or library from the same individual, to evaluate its effect on the molecular complexity of the experiment. We validated our experiments with 18 chimpanzee faecal samples collected from two field sites as a part of the Pan African Programme: The Cultured Chimpanzee. Those two field sites are in Kibale National Park, Uganda (N = 9) and Loango National Park, Gabon (N = 9). We demonstrate that at least 16 libraries can be pooled, target enriched through hybridization, and sequenced allowing for the genotyping of 951,949 exome markers for population genetic analyses. Further, we observe that molecule richness, and thus, data acquisition, increase when using multiple libraries from the same extract or multiple extracts from the same sample. Finally, repeated captures significantly decrease the proportion of off-target reads from 34.15% after one capture round to 7.83% after two capture rounds, supporting our conclusion that two rounds of target enrichment are advisable when using complex faecal samples.
AB - Target-capture approach has improved over the past years, proving to be very efficient tool for selectively sequencing genetic regions of interest. These methods have also allowed the use of noninvasive samples such as faeces (characterized by their low quantity and quality of endogenous DNA) to be used in conservation genomic, evolution and population genetic studies. Here we aim to test different protocols and strategies for exome capture using the Roche SeqCap EZ Developer kit (57.5 Mb). First, we captured a complex pool of DNA libraries. Second, we assessed the influence of using more than one faecal sample, extract and/or library from the same individual, to evaluate its effect on the molecular complexity of the experiment. We validated our experiments with 18 chimpanzee faecal samples collected from two field sites as a part of the Pan African Programme: The Cultured Chimpanzee. Those two field sites are in Kibale National Park, Uganda (N = 9) and Loango National Park, Gabon (N = 9). We demonstrate that at least 16 libraries can be pooled, target enriched through hybridization, and sequenced allowing for the genotyping of 951,949 exome markers for population genetic analyses. Further, we observe that molecule richness, and thus, data acquisition, increase when using multiple libraries from the same extract or multiple extracts from the same sample. Finally, repeated captures significantly decrease the proportion of off-target reads from 34.15% after one capture round to 7.83% after two capture rounds, supporting our conclusion that two rounds of target enrichment are advisable when using complex faecal samples.
KW - conservation genetics
KW - exome
KW - next-generation sequencing
KW - noninvasive samples
KW - target enrichment
UR - http://www.scopus.com/inward/record.url?scp=85037375358&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85037375358&partnerID=8YFLogxK
U2 - 10.1111/1755-0998.12728
DO - 10.1111/1755-0998.12728
M3 - Article
C2 - 29058768
AN - SCOPUS:85037375358
SN - 1755-098X
VL - 18
SP - 319
EP - 333
JO - Molecular Ecology Resources
JF - Molecular Ecology Resources
IS - 2
ER -