TaqMan Multiplex qPCR Method to Genotype PARG Knockout Mice.

Abstract

Members of PARP family are responsible for poly(ADP-ribose) (pADPr) posttranslational modification synthesis. They are intensively studied proteins with more than 20,500 related papers in PubMed database search to date. PARG, the main enzyme that degrades pADPr, is unfairly attracted less attention, and 40 times less papers (a little more than 500) are related to its functioning. The difficulties to work with PARG knockout animals due to its early embryo lethality could be one reason for this huge difference. Mice PARG-specific antibodies are not available from any vendor, which also complicates the research process. There is one available for public PARG knockout mice line generated by KOMP project. It has LacZ cassette, which replaces three critical exons in PARG gene. Here, we present the method to genotype these mice with Taqman qPCR multiplex approach. It allows to work with a small amount of DNA material like early embryo stages and to separate maternal DNA contamination. The modification of this method is also applicable for studying PARG conditional knockouts and identifying the success of floxed PARG gene exon deletion by Cre-driven recombination.