Comparing Genotyping Accuracy Using Buccal Swabs versus Tail Biopsies by PCR in B6;C3-Tg(Prnp-SNCA*A53T)83Vle and B6;C3-Tg(Prnp-SNCA*A53T)83Vle Snca tm1Mjff Mice.

Autor: Lui MF; Institute for Neurodegenerative Diseases, University of California San Francisco, San Francisco, California., Osborne M; GenoTyping Center of America, Waterville, Maine., Dehm T; GenoTyping Center of America, Waterville, Maine., Lee M; GenoTyping Center of America, Waterville, Maine., Castaneda JA; Institute for Neurodegenerative Diseases, University of California San Francisco, San Francisco, California.
Jazyk: angličtina
Zdroj: Journal of the American Association for Laboratory Animal Science : JAALAS [J Am Assoc Lab Anim Sci] 2024 Jul 01; Vol. 63 (4), pp. 363-367. Date of Electronic Publication: 2024 Apr 22.
DOI: 10.30802/AALAS-JAALAS-23-000045
Abstrakt: Genotyping is a common and necessary procedure performed on genetically modified animals to distinguish carriers from noncarriers of the variants of interest. Established methods involve collection of tissues such as tips of tails or notches of ears. Noninvasive methods have been described but not widely adopted for reasons including inertia to change, needs to adjust PCR protocols, and the lack of validation; noninvasive genotyping methods are a refinement on animal welfare, but questions remain regarding how they compare with invasive methods in terms of genotyping accuracy rate and reproducibility. To gain answers to these questions, we compared the detection accuracy of the transgene and determination of zygosity in B6;C3-Tg(Prnp-SNCA*A53T)83Vle and B6;C3-Tg(Prnp-SNCA*A53T)83Vle Snca tm1Mjff neonatal mice between tail biopsies and buccal swabs. Moreover, we weighed and observed mice following genotyping to see if any clinical differences can be discerned. Weight data did not support statistically significant differences in mice undergoing different genotyping procedures and control. No statistically significant difference was found between using buccal swabs or tail biopsies for genotyping with PCR or quantitative PCR. None of the pups swabbed was rejected by the dam. Our findings indicate that buccal swabbing is a more humane and feasible alternative to tail biopsies for high-throughput genotyping.
Databáze: MEDLINE