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Evaluation of a reliable and cost-effective method of DNA isolation for mouse genotyping

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Abstract

Genotyping is commonly used to define specific gene alterations or the presence of transgenes in mice. This procedure is typically done using DNA isolated from mouse tail tissue. Although there are commercially available kits for tail DNA isolation, they can be time consuming and costly for routine genotyping. In this study, we describe a rapid, “crude” DNA isolation method using mouse tail tissue and compare it to a frequently used, commercially available kit in the genotyping of over 1,000 total mice from 8 genetic lines. Our genotyping results were obtained faster and less expensively but with the same success rate (Crude method: 97.7 %, Kit method: 98.4 %). To our knowledge, this is the first systematic study to compare the reliability of this crude DNA isolation method for mouse genotyping compared to a commercially available kit.

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Acknowledgments

This study was supported by the Northwestern Faculty Foundation Dixon Innovation Award (Northwestern University, Chicago, IL) and the Pfizer ASPIRE (Advancing Science through Pfizer—Investigator Research Exchange) Award (Pfizer Inc.), both awarded to Dr. Jiwang Chen. We thank Dr. Youyang Zhao at the University of Illinois at Chicago for generously providing the eNOS knockout mice and primers used in this study.

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Authors and Affiliations

  1. Institute for Personalized Respiratory Medicine, Section of Pulmonary, Critical Care, Sleep and Allergy, Department of Medicine, University of Illinois at Chicago, Chicago, IL, 60612, USA

    Justin R. Sysol, Carrie Kempf, Michelle N. Helton, Yangbasai Dong, Joe G. N. Garcia, Roberto F. Machado & Jiwang Chen

  2. Department of Medicine, Feinberg School of Medicine, Northwestern University, Chicago, IL, 60611, USA

    Daocheng Zhu, Haiying Sun & Jiwang Chen

Authors
  1. Justin R. Sysol
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  2. Carrie Kempf
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  3. Michelle N. Helton
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  4. Yangbasai Dong
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  5. Daocheng Zhu
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  6. Haiying Sun
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  7. Joe G. N. Garcia
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  8. Roberto F. Machado
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  9. Jiwang Chen
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Correspondence to Jiwang Chen.

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Sysol, J.R., Kempf, C., Helton, M.N. et al. Evaluation of a reliable and cost-effective method of DNA isolation for mouse genotyping. Biotechnol Lett 35, 509–514 (2013). https://doi.org/10.1007/s10529-012-1113-2

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  • DOI: https://doi.org/10.1007/s10529-012-1113-2

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