Well. I have some transgenic mice. And also have their genomic DNA. Now, I would like to identify that which one is homozygous and heterozygous. I don't want to collect sample from mice to extract RNA, to produce cDNA and qPCR. Its a lengthy process. So, in this situation, which one is the cheapest and quickest way to identify the mice and how?
You don't need to go the whole RNA-cDNA-qPCR route; rather, you can do qPCR directly on the genomic DNA samples you already have. How to design the primers depends on the specific mutation that was introduced.
Hope it helps!
Edit: you don't even need qPCR. Conventional PCR will do if the design is right.
You don't need to use qPCR at all. Just design PCR primers that will differentiate the two alleles. The exact method will depend on the difference between the wildtype and transgenic alleles. Talk to whoever made these mice. They must have used some method to genotype their mice during the breeding process. Good luck!
Do you know the location of the transgene? Is it in a specific locus, such as Rosa26? Then, you should know the flanking region of the transgene for design PCR primer pairs.
Also, if there is a lab around you familiar with qPCR, they probably can help to run it with you and get the answer quickly.
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