When crossing loxP mice with FLPe mice, LacZ and Neo are removed; how do you perform genotyping in this case? Additionally, can you determine if a loxP mouse is heterozygous or homozygous through genotyping?
When crossing loxP mice with FLPe mice, the removal of LacZ and Neo leaves behind the loxP site. To genotype these mice, you can use the following methods:
As for determining heterozygosity or homozygosity:
- Heterozygous: If you observe both the wt and loxP bands, the mouse is heterozygous (wt/loxP).
- Homozygous: If only the loxP band is present, the mouse is homozygous (loxP/loxP).
Remember to validate your genotyping results and adjust primer design as needed.
I am attaching some articles for your reference-
Article PCR-Based Strategy for Genotyping Mice and ES Cells Harborin...
Article One-Step Genotyping Method in loxP-Based Conditional Knockou...
https://www.nature.com/articles/ng0600_139.pdf
https://www.jax.org/news-and-insights/2006/may/the-cre-lox-and-flp-frt-systems
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