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Non‐invasive transgenic mouse genotyping using stool analysis
Author(s) -
Broome Rosemary L.,
Feng Li,
Zhou Qin,
Smith Alicia,
Hahn Nina,
Matsui Suzanne M.,
Omary M.Bishr
Publication year - 1999
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(99)01516-1
Subject(s) - genotyping , transgene , biology , genetically modified mouse , polymerase chain reaction , microbiology and biotechnology , gene , real time polymerase chain reaction , genotype , genetics
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.