Hi, I have to genotype nestin-GFP mice. But I've found that there is only available a PCR reaction that only can tells if the construction is present in the mouse, but can not distinguish between homocigotic or heterocigotic.
¿anyone have faced this problem and have a solution?¿primers to amplify the wt allele if present?¿maybe a qPCR experiment to disthinguish homo or heterocigotic?
Nestin-GFP mice are to my knowledge transgenic mice with GFP under the control of the Nestin promoter, so it doesn´t matter whether the animals are hetero- or homozygote because GFP will be expressed independent of the nestin wild type. It´s only important for breeding and in this case nestin primers won´t help you very much.
Thanks a lot!. The problem come from this point: we found problems to keep the line, sometimes you only have wt... and there is other critical point: bibliography tells that homozygote can carry health problems with the time. This is my concern.
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