I am trying to breed two types of Rosa26 knock-in mice together, but I have not had a single litter from 2 breeding pairs over 3 months. The breeders are now 6 months old. One breeder is a tdTomato reporter mouse (Ai14): Rosa26tdTomato/tdTomato. The other is the Ai87 mouse: Rosa26iGluSnFr/iGluSnFr. Technically, I believe the whole insert into the Rosa26 locus is Rosa26 - CAGpromoter - LoxP/stop/LoxP - then the gene sequence (tdTomato or iGluSnFr). I naively thought the offspring would all be Rosa26tdTomato/iGluSnFr and the alleles would segregate that way. Anyone have some molecular genetics insights? or could this just be bad luck?
Have you seen any mating behavior or evidence of pregnancy (mucous plug)? Got to rule out the obvious before we get to the possibilities of weird genetics.
If both breeders are pure line (or homozygous for the alleles), you should have the genotype you want in the offspring. How many litters have you analyzed? How did you test the co-exist of the two alleles in the offspring, PCR?
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