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Open Access Research article

A systematic evaluation of hybridization-based mouse exome capture system

Qingsong Gao, Wei Sun, Xintian You, Sebastian Froehler and Wei Chen*

Author Affiliations

Laboratory for Novel Sequencing Technology, Functional and Medical Genomics, Berlin Institute for Medical Systems Biology, Max-Delbrück-Centrum für Molekulare Medizin, Robert-Rössle-Straße 10, Berlin 13125, Germany

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BMC Genomics 2013, 14:492  doi:10.1186/1471-2164-14-492

Published: 21 July 2013

Abstract

Background

Exome sequencing is increasingly used to search for phenotypically-relevant sequence variants in the mouse genome. All of the current hybridization-based mouse exome capture systems are designed based on the genome reference sequences of the C57BL/6 J strain. Given that the substantial sequence divergence exists between C57BL/6 J and other distantly-related strains, the impact of sequence divergence on the efficiency of such capture systems needs to be systematically evaluated before they can be widely applied to the study of those strains.

Results

Using the Agilent SureSelect mouse exome capture system, we performed exome sequencing on F1 generation hybrid mice that were derived by crossing two divergent strains, C57BL/6 J and SPRET/EiJ. Our results showed that the C57BL/6 J-based probes captured the sequences derived from C57BL/6 J alleles more efficiently and that the bias was higher for the target regions with greater sequence divergence. At low sequencing depths, the bias also affected the efficiency of variant detection. However, the effects became negligible when sufficient sequencing depth was achieved.

Conclusion

Sufficient sequence depth needs to be planned to match the sequence divergence between C57BL/6 J and the strain to be studied, when the C57BL/6 J–based Agilent SureSelect exome capture system is to be used.

Keywords:
Mouse exome capture system; Sequence divergence; Capture bias; Efficiency of variant detection