Mice Genotyping Using Buccal Swab Samples: An Improved Method |
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Authors: | Dinko Mitrečić Sandra Mavrić Božica Vrabec Branica Srećko Gajović |
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Institution: | (1) Croatian Institute for Brain Research, School of Medicine, University of Zagreb, Šalata 12, HR-10000 Zagreb, Croatia |
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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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Keywords: | DNA isolation Buccal swabs Genotyping PCR Mouse |
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