Cfx96 touch real time pcr detection system machine

Mice genetically deficient in Klk6, or wild type litter mates, were used in all in vivo experiments. Both male and female mice were used and ranged in age from postnatal day (P) 0 to 90 days. Coding exons 3 and 4 of the Klk6 gene were deleted in knockouts by inserting a “universal docking site” cassette (Samantha Sykioti et al. 2020 ) (Dr. A. Nagy, Mount Sinai, Toronto, Canada) and were a generous gift from Dr. G. Sotiropoulou, University of Patras, Greece. A schematic diagram of the transgenic cassette and the PCR genotyping strategy were recently published (Pampalakis et al. 2017 (link)). Mice with Klk6 knockout are viable, fertile and have no prominent phenotypic abnormalities. Wild-type (Klk6+/+) littermates were used as controls for all experiments. Mice were genotyped using probes and primers as listed in Table 1, using iTaq™ Universal Probes One-Step Kit or the iTaq™ Universal SYBR Green One-Step Kit (1725141 or 1725151, respectively, Bio-Rad) on a CFX96 Touch Real-Time PCR Detection system machine (Bio-Rad). Klk6 knockout, Klk6 wild type and C57BL/6J mice (#000664, Jackson Laboratory Bar Harbor, ME) were used for the study of Klk6 in primary oligodendrocyte progenitor cell cultures. All animal experiments were carried out with adherence to NIH Guidelines for animal care and safety and were approved by the Mayo Clinic Institutional Animal Care and Use Committee.