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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

Biochemical Genetics

|January 24, 2008

Summary

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.

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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.

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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.