Identification of transgenic mice by direct PCR analysis of lysates of epithelial cells obtained from the inner surface of the rectum.
Conventional screening protocols for transgene integration in mice employ tail tips or blood samples as sources to obtain genomic DNA preparations. We have developed a simple alternative non-surgical method. Epithelial cells are scraped off the inner surface of the rectum with a sterile plastic inoculation loop and are lysed with Kawasaki buffer. The lysate can be directly examined in a polymerase chain reaction (PCR) analysis without any need for further DNA purification. This procedure causes minimal harm and stress to the animals and repeated samples can be obtained as often as necessary. This technique has been used successfully to identify transgenic mice from a number of different lines. The method allows quick screening of numerous animals and contributes to a reduction of the number of surgical biopsies required.
Lahm
H
H
Institute of Molecular Animal Breeding, Genecenter, Munich, Germany.
Hoeflich
A
A
Rieger
N
N
Wanke
R
R
Wolf
E
E
D000818:Animals; D018076:DNA, Complementary / Q000032:analysis; D004847:Epithelial Cells; D051379:Mice; D008822:Mice, Transgenic; D016133:Polymerase Chain Reaction / Q000379:methods*; D012007:Rectum / Q000166:cytology*; D019076:Transgenes
D018076:DNA, Complementary