Rapid genotyping of newborn gene mutant mice |
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Authors: | Henneberger C Grantyn R Rothe T |
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Affiliation: | Developmental Physiology, Johannes Müller Institute for Physiology, Charité, Tucholskystr. 2, D-10117 Berlin, Germany |
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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. |
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Keywords: | Genomic DNA isolation PCR Brain-derived neurotrophic factor (BDNF) Neurotrophin mutants Knockout mice |
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