Rapid genotyping of newborn gene mutant mice.
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.
Henneberger
C
C
Developmental Physiology, Johannes Müller Institute for Physiology, Charité, Tucholskystr. 2, D-10117, Berlin, Germany.
Grantyn
R
R
Rothe
T
T
D000818:Animals; D000831:Animals, Newborn / Q000235:genetics*; D019208:Brain-Derived Neurotrophic Factor / Q000172:deficiency / Q000235:genetics; D004252:DNA Mutational Analysis / Q000379:methods*; D005820:Genetic Testing / Q000379:methods*; D005838:Genotype; D051379:Mice; D018345:Mice, Knockout / Q000235:genetics*; D016133:Polymerase Chain Reaction; D013997:Time Factors
D019208:Brain-Derived Neurotrophic Factor