通过 PCR 比较 B6;C3-Tg(Prnp-SNCA*A53T)83Vle 和 B6;C3-Tg(Prnp-SNCA*A53T)83Vle Sncatm1Mjff 小鼠口腔拭子与尾部活检的基因分型准确性。

Ming F Lui Dvm, Melissa Osborne Ms, Todd Dehm Ma, Min Lee Ba, Julian A Castaneda Dvm PhD Daclam
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引用次数: 0

摘要

基因分型是对转基因动物进行基因分型的一种常见且必要的程序,用于区分相关变异体的携带者和非携带者。现有的方法需要采集组织,如尾巴尖或耳朵的缺口。无创基因分型方法是对动物福利的一种改进,但在基因分型准确率和可重复性方面与有创方法的比较仍存在问题。为了回答这些问题,我们比较了 B6;C3-Tg(Prnp-SNCA*A53T)83Vle和 B6;C3-Tg(Prnp-SNCA*A53T)83Vle Sncatm1Mjff 新生小鼠尾部活检与颊拭子之间的转基因检测准确性和子系测定。此外,我们还对基因分型后的小鼠进行了称重和观察,以确定是否存在临床差异。不同基因分型程序和对照组小鼠的体重数据在统计学上没有显著差异。使用口腔拭子或尾部活检进行 PCR 或定量 PCR 基因分型在统计学上没有明显差异。没有一只幼鼠被母鼠排斥。我们的研究结果表明,在进行高通量基因分型时,颊拭子比尾巴活检更人道、更可行。
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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 Sncatm1Mjff Mice.
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 Sncatm1Mjff 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.
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American Society of Laboratory Animal Practitioners Position Statement: Handling and Physical Restraint of Research Animals. American Society of Laboratory Animal Practitioners Position Statement: Definition of Animal Welfare. Effect of Novel High-fat Diet Feeding Methods on Food Wastage, Weight Gain, Hair Coat Grease Accumulation, and Scratching Behavior in C57BL/6NCrl Mice. Identification and Treatment of Fur Mites (Radfordia lemnina) in California Deer Mice (Peromyscus californicus) Using Selamectin. American Society of Laboratory Animal Practitioners Position Statement: Animal Care Principles.
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