DNA detection in hair of transgenic mice--a simple technique minimizing the distress on the animals.
The breeding of transgenic animals requires that each individual offspring be analysed for integration of transgenic deoxyribonucleic acid (DNA), unless exclusively homozygous animals are mated. The standard protocol for identification of transgenic animals (Hogan et al. 1994) is based on tissue samples and preparation of chromosomal DNA including proteinase K digestion and phenol/chloroform extraction. The procedure described here represents a much simpler and faster method to screen offspring for the transgene DNA. It is based on the use of hair bulbs as sample material, which can be directly used for polymerase chain reaction (PCR) after alkaline lysis. This protocol allows large numbers of animals to be easily screened in a minimum amount of time. A unique advantage though, is the reduction of the distress caused to the animals. With respect to the 3Rs (Replacement, Reduction, Refinement), and because of technical advantages this method may replace ear or tail clipping.
Schmitteckert
E M
EM
Institute of Pharmacology and Toxicology, University of Wuerzburg, Germany.
Prokop
C M
CM
Hedrich
H J
HJ
D000827:Animal Welfare; D000818:Animals; D004247:DNA / Q000032:analysis*; D017931:DNA Primers / Q000737:chemistry; D005821:Genetic Techniques; D006197:Hair / Q000737:chemistry*; D051379:Mice; D008822:Mice, Transgenic / Q000235:genetics; D016133:Polymerase Chain Reaction; D013623:Tail / Q000737:chemistry / Q000293:injuries
D017931:DNA Primers; D004247:DNA