Hello,
I am using a mice line which is a cross product of Sun1-tagged mice (Jax #021039) with VIP-Cre mice (Jax # 010908).
I'm trying to establish a PCR genotyping protocol to detect whether an unintended recombination occurred at some point during the embryonic development.
Since I couldn't find a map for the KI sequence in the Sun1-tagged mouse - the process of planning the primers for PCR genotyping is a bit tricky.
I would appreciate if someone that has performed this type of genotyping for the ROSA26-pCAG-LSL allele could help me with this.
Alternatively, it would be also very helpful if someone could refer me to where I can find a map for Jax #021039 mice. I couldn't find it through MGI (or perhaps I missed it).
Many thanks!
Eran
Hi Eran,
Jackson Laboratories provides primer sequences for genotyping their mouse lines that can differentiate between WT, mutant, and heterozygous.
Stock No: 010908
https://www.jax.org/Protocol?stockNumber=010908&protocolID=29387
Stock No: 021039
https://www.jax.org/Protocol?stockNumber=021039&protocolID=31117
Or you are looking for information to genotype some other loci?
Best,
Sheng
Thank you Sheng Sun ;
You right, Jackson indeed provides the genotyping protocol & primers to determine whether or not you have the mutant allele.
However, what I'm missing is the primers to detect whether or not a recombination occurred. If I'd used the WT vs. mutant protocol I'll get a positive result for a mutant allele regardless the presence or absence of a Cre-mediated recombination.