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Non-invasive genotyping of transgenic animals using fecal DNA

Lab Animal volume 41, pages 102–107 (2012)Cite this article

Abstract

For genotyping of transgenic animals, many IACUC guidelines recommend the use of fecal DNA when possible because this approach is non-invasive. Existing methods for extracting fecal DNA may be costly or involve the use of toxic organic solvents. Furthermore, feces contain an abundance of PCR inhibitors that may hinder DNA amplification when they are co-purified with fecal DNA. Here the authors describe a cost-effective, non-toxic method for genotyping transgenic animals by using the reagent AquaStool to extract fecal DNA and remove PCR inhibitors. Genotyping results obtained from fecal DNA samples extracted using AquaStool were reliably accurate when compared with results obtained from tail DNA samples. Because it is non-invasive, the authors believe that use of this method for genotyping transgenic animals using fecal DNA samples may improve animal welfare.

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Figure 1: Genotyping results of ten adult mice from the pKZ1 transgenic line using fecal DNA samples prepared using the AquaStool reagent.
Figure 2: Genotyping results of five Zp3-cre transgenic mice (59, 61, 68, 70 and 71) using fecal DNA samples shipped after being preserved in AquaStool solution or air-dried overnight.
Figure 3: PCR amplification of a 370-bp sequence from mouse chromosome 6 (Chr. 6) from AquaStool-prepared DNA with or without centrifugation to remove water-insoluble material that included PCR inhibitors.

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Acknowledgements

This work was funded in part by a grant from the Johns Hopkins Center for Alternatives to Animal Testing (CAAT).

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Authors and Affiliations

  1. MultiTarget Pharmaceuticals LLC, Salt Lake City, UT

    Zhidong Chen

  2. Atomic Energy of Canada Ltd., Chalk River Laboratories, Chalk River, Canada

    Rebecca R. Mantha

  3. Department of Biology, Johns Hopkins University, Baltimore, MD

    Janice S. Chen

  4. Aab Cardiovascular Research Institute, University of Rochester School of Medicine and Dentistry, Rochester, NY

    Orazio J. Slivano

  5. Genetic Engineering Core, National Eye Institute, National Institutes of Health, Bethesda, MD

    Hideko Takahashi

Authors
  1. Zhidong Chen
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  2. Rebecca R. Mantha
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  3. Janice S. Chen
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Corresponding author

Correspondence to Zhidong Chen.

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Competing interests

The authors declare competing financial interests. Z.C. is an employee of MultiTarget Pharmaceuticals, LLC, which is the manufacturer of AquaStool.

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Chen, Z., Mantha, R., Chen, J. et al. Non-invasive genotyping of transgenic animals using fecal DNA. Lab Anim 41, 102–107 (2012). https://doi.org/10.1038/laban0412-102

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