A simplified method to prepare PCR template DNA for screening of transgenic and knockout mice.
Polymerase chain reaction (PCR) amplification of DNA is the most widely used technique for screening of large numbers of genetically engineered transgenic or knockout mice (Mus musculus). In this report, we present a new DNA preparation procedure for running diagnostic PCR. In this procedure, mouse ear tissue was used directly for PCR after the tissue underwent brief digestion in a solution containing only proteinase K. Using this method, we have successfully screened several lines of single, double, and triple transgenic and knockout mice. The results are reliable and reproducible. The advantage of this new method is that DNA purification by organic extraction or isolation kit was omitted. DNA purification is the limiting factor in terms of time and money when screening transgenic and knockout mice by PCR. In addition, using ear instead of tail tissue can reduce distress of animals because the samples can be obtained when the mice are labeled by ear punch.
Ren
S
S
Institute of Human Virology, Division of Infectious Diseases, Department of Medicine and the University of Maryland Medical School, Baltimore, Maryland 21201, USA.
Li
M
M
Cai
H
H
Hudgins
S
S
Furth
P A
PA
D000823:Animal Identification Systems; D000818:Animals; D004247:DNA / Q000032:analysis*; D004431:Ear, External / Q000737:chemistry; D005820:Genetic Testing / Q000379:methods*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout / Q000235:genetics*; D008822:Mice, Transgenic / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D013698:Templates, Genetic
D004247:DNA