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Sci Rep. 2015 Aug 3;5:12810. doi: 10.1038/srep12810.

Improved canine exome designs, featuring ncRNAs and increased coverage of protein coding genes.

Author information

1
Laboratory of Pharmaceutical Biotechnology, Faculty of Pharmaceutical Sciences, Ghent University, Ghent, Belgium.
2
Institut de Génétique et Développement de Rennes, CNRS-URM6290, Université Rennes1, Rennes, France.
3
Department of Applied Biosciences, Faculty of Bioscience Engineering, University College Ghent, Ghent, Belgium.
4
Department of Medical Imaging and Small Animal Orthopaedics, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
5
Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
6
1] Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA [2] Science for Life Laboratory, Department of Medical Biochemistry and Microbiology, Uppsala University, Uppsala, Sweden.
7
Department of Medicine and Clinical Biology of Small Animals, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.

Abstract

By limiting sequencing to those sequences transcribed as mRNA, whole exome sequencing is a cost-efficient technique often used in disease-association studies. We developed two target enrichment designs based on the recently released annotation of the canine genome: the exome-plus design and the exome-CDS design. The exome-plus design combines the exons of the CanFam 3.1 Ensembl annotation, more recently discovered protein-coding exons and a variety of non-coding RNA regions (microRNAs, long non-coding RNAs and antisense transcripts), leading to a total size of ≈ 152 Mb. The exome-CDS was designed as a subset of the exome-plus by omitting all 3' and 5' untranslated regions. This reduced the size of the exome-CDS to ≈ 71 Mb. To test the capturing performance, four exome-plus captures were sequenced on a NextSeq 500 with each capture containing four pre-capture pooled, barcoded samples. At an average sequencing depth of 68.3x, 80% of the regions and well over 90% of the targeted base pairs were completely covered at least 5 times with high reproducibility. Based on the performance of the exome-plus, we estimated the performance of the exome-CDS. Overall, these designs provide flexible solutions for a variety of research questions and are likely to be reliable tools in disease studies.

PMID:
26235384
PMCID:
PMC4522663
DOI:
10.1038/srep12810
[Indexed for MEDLINE]
Free PMC Article

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