Hello,
I have recently begun to introduce the Brainbow 2.1 gene to some of our inducible Cre mice. We have used the mT/mG reporter in the past with success. I recently had some extra master mix while setting up a reaction for mT/mG and ran the reaction using DNA from heterozygous and homozygous Brainbow 2.1 mice.
To my surprise bands appeared in the reaction as if they were heterozygous and homozygous for mT/mG.
The strains used are from Jax as follows:
https://www.jax.org/strain/007576
https://www.jax.org/strain/013731
Is there some type of overlap in the transgene construct that is resulting in bands from the Brainbow mice DNA in the mT/mG reaction?
Any input would be appreciated!
Thanks
Kyle
Two related likely answers:
When you're genotyping for the mt/mg allele, you're probably just checking for tdTomato or GFP. For the wild type allele, you're likely probing for the intact Rosa26 gene.
The Brainbow allele is in the same R26 PSor locus, and contains the same general fluorescent proteins as the mt/mg one. As such, you're able to use the generic probes you were already using for mt/mg genotyping.
In other words, you're likely genotyping/probing for "fluorescent reporter" and for "intact Rosa26 locus".
I'm fairly certain this is the case, but we'd have to know your primers to be certain!
Hope this clears it up.
Best,
Alberto