Repozitorij Medicinskog fakulteta Sveučilišta u Zagrebu

Mice genotyping using buccal swab samples: an improved method

Mitrečić, Dinko and Mavrić, Sandra and Vrabec Branica, Božica and Gajović, Srećko (2008) Mice genotyping using buccal swab samples: an improved method. Biochemical Genetics, 46 (3-4). pp. 105-12. ISSN 0006-2928

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    English 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.

    Item Type: Article
    MeSH: Animals ; Cheek ; DNA/isolation & purification ; Genotype ; Mice ; Mouth Mucosa ; Polymerase Chain Reaction ; Time Factors
    Divisions: Katedra za histologiju i embriologiju
    Hrvatski institut za istraživanje mozga
    Depositing User: Marijan Šember
    Status: Published
    Mitrečić, Dinko
    Mavrić, Sandra
    Vrabec Branica, Božica
    Gajović, Srećko
    Date: April 2008
    Date Deposited: 25 Feb 2010
    Last Modified: 23 Sep 2011 18:11
    Subjects: /
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