Wt1CREERT2-RosaYFP Model... Is it possible to asses recombination through PCR (DNA)?

21 April 2021 1 332 Report

Hello,

As the tittle says... Since tamoxifen induces DNA recombination, is it possible to detect this recombination using specific primers? I am also assessing recombination through immunohistochemistry, but I was looking for other methods.

Thanks

Cristina

Lahiru Chamara Weerasinghe Arachchige

Yes, you can. You need to remember that WT1 is not expressed in all cells. If you do not isolate YFP positive cells, you will have some contaminations from other cells. You can FACS your cell before genotyping to avoid the problem.

Best way to do a qPCR. You can check the level of expression (at cDNA level) for your gene of interest, and compare the expression level with your control groups (Cre without TAM induction, Flox controls and etc).

Good luck

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