Gene targeting in mouse ES cells replaces or modifies genes of interest; conditional alleles, reporter knock-ins, and amino acid changes are common examples of how gene targeting is used. To streamline and increase the efficiency in our ES cell pipeline and decrease the timeline for mouse models produced via ES cells, automation is introduced in the pipeline. Below, we describe a novel and effective approach utilizing ddPCR, dPCR, automated DNA purification, MultiMACS, and adenovirus recombinase combined screening workflow that reduces the time between therapeutic target identification and experimental validation.
Keywords: Biomek i5; Genotyping; Mouse embryonic stem cells (ESC); MultiMACS (magnetic-activated cell sorting); dPCR (digital PCR); ddPCR (droplet digital PCR).
© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.