Abstract
Routine methods used to genotype mice involve isolation of DNA from partially amputated neonate’s tail, toe, or ear. The inevitable drawbacks of such techniques are the animal’s pain response and the increased time and funds required for DNA purification. In order to implement a noninvasive and simple protocol for mouse DNA isolation, we have improved the method based on samples collected by swabbing of the inner cheek. Combining alkaline and temperature lysis, it was possible to isolate a DNA solution ready for PCR in less than an hour. Testing the method on three different mouse lines showed that it is highly efficient, the volume of the PCR samples could be reduced to 25 μl, and fragments up to 800 bp were successfully amplified. This protocol reduces animal discomfort, shortens the time for DNA isolation, and enables amplification of larger DNA fragments with optimal success rate, thus considerably facilitating large-scale genotyping of different mouse lines.
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Acknowledgments
This work was supported by grants from the Ministry of Science, Education and Sports, Republic of Croatia (No. 108-1081870-1902) and International Center for Genetic Engineering and Biotechnology (No. CRP/CRO06-02). We thank Achim Gossler for the Not eGFP mouse line, Bojan Macanović for comments regarding the English in the manuscript, and Ljiljana Krznar for animal care.
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Mitrečić, D., Mavrić, S., Branica, B.V. et al. Mice Genotyping Using Buccal Swab Samples: An Improved Method. Biochem Genet 46, 105–112 (2008). https://doi.org/10.1007/s10528-007-9133-7
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DOI: https://doi.org/10.1007/s10528-007-9133-7