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Mice genotyping using buccal swab samples: an improved method
Dinko Mitrecić1, Sandra Mavrić, Bozica Vrabec Branica
1Croatian Institute for Brain Research, School of Medicine, University of Zagreb, Salata 12, HR-10000 Zagreb, Croatia. dominic@mef.hr
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 microl, 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.
Insights
This study introduces a noninvasive cheek swab method for mouse DNA isolation, significantly reducing animal distress and processing time. The improved protocol yields DNA suitable for polymerase chain reaction (PCR) amplification, facilitating large-scale mouse genotyping.
Area of Science:
- Animal research methodologies
- Molecular biology techniques
- Genetics and genomics
Background:
- Traditional mouse DNA isolation methods involve tail, toe, or ear amputation, causing animal pain and requiring extensive DNA purification.
- These conventional techniques are time-consuming and costly, posing challenges for large-scale genetic studies.
Purpose of the Study:
- To develop a noninvasive and simplified protocol for mouse DNA isolation.
- To reduce animal discomfort and laboratory resource utilization in mouse genotyping.
Main Methods:
- Improved DNA isolation using cheek swabs instead of tissue amputation.
- Combined alkaline and temperature lysis for rapid DNA extraction.
- Utilized polymerase chain reaction (PCR) for DNA amplification and analysis.
Main Results:
- Isolated DNA solution ready for PCR in under an hour.
- Demonstrated high efficiency across three different mouse lines.
- Successfully amplified DNA fragments up to 800 base pairs (bp) using reduced PCR sample volumes (25 microliters).
Conclusions:
- The noninvasive cheek swab method significantly reduces animal pain and isolation time.
- This protocol is highly efficient for large-scale mouse genotyping.
- Enables successful amplification of larger DNA fragments, facilitating genetic studies.
