Mice genotyping using buccal swab samples: an improved method.
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.
Mitrecić
Dinko
D
Croatian Institute for Brain Research, School of Medicine, University of Zagreb, Salata 12, HR-10000 Zagreb, Croatia. dominic@mef.hr
Mavrić
Sandra
S
Branica
Bozica Vrabec
BV
Gajović
Srećko
S
D000818:Animals; D002610:Cheek; D004247:DNA / Q000302:isolation & purification*; D005838:Genotype; D051379:Mice; D009061:Mouth Mucosa; D016133:Polymerase Chain Reaction; D013997:Time Factors
D004247:DNA