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Rapid genotyping of newborn gene mutant mice
Authors:Henneberger C  Grantyn R  Rothe T
Affiliation:

Developmental Physiology, Johannes Müller Institute for Physiology, Charité, Tucholskystr. 2, D-10117 Berlin, Germany

Abstract:One important aspect of utilizing transgenic mice is the need to genotype them in order to distinguish mice that carry a disrupted gene or a transgene from mice that do not. Current methods for genotyping include isolation of genomic DNA from tail biopsies followed by PCR amplification. Particularly, both digestion of tail tissue using proteinase K as well as resuspension of purified DNA are time-consuming and were usually carried out overnight. Here, we describe a rapid and robust method for the genotyping of bdnf targeted mice which allows us to determine the genotype of newborn mice at the day of birth within 6 h. After a freezing–thawing step tail tissue is digested in less than 2 h, and the DNA is precipitated, resuspended and ready for PCR in about 60 min. The method could be easily adapted to a variety of different mutant mice and especially should benefit neuroscientists interested in using animals with known genotype very early in postnatal development.
Keywords:Genomic DNA isolation   PCR   Brain-derived neurotrophic factor (BDNF)   Neurotrophin mutants   Knockout mice
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