Non-invasive transgenic mouse genotyping using stool analysis.
Commonly applied genotyping of transgenic mice involves using tail or ear biopsies which may cause discomfort to the animal. We tested the possibility of polymerase chain reaction (PCR)-based mouse genotyping using stool specimens from three transgenic mouse lines that overexpress 10-18 transgene copies of human keratin polypeptide 18, as compared to genotyping using tail biopsies. Stool specimens were obtained with ease and provided easy detection of the human transgene product. The method was also able to detect endogenous mouse actin and keratin genes which presumably are present at two copies each. Nested PCR was not necessary for genotyping using stool-derived genomic material but did increase the relative magnitude of the signal obtained. The non-invasive genotyping method described herein offers a reproducible, sensitive and effective modality that could replace invasive tissue sampling procedures currently used to test thousands of genetically altered mice.
Broome
R L
RL
Department of Veterinary Medicine, VA Palo Alto Health Care System, 3801 Miranda Avenue, 154J, Palo Alto, CA 94304, USA.
Feng
L
L
Zhou
Q
Q
Smith
A
A
Hahn
N
N
Matsui
S M
SM
Omary
M B
MB
D000818:Animals; D001706:Biopsy / Q000379:methods; D005243:Feces / Q000737:chemistry*; D005838:Genotype; D006801:Humans; D007633:Keratins / Q000032:analysis / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*
D007633:Keratins