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Updated: Oct 26, 2025

Using a Fluorescent PCR-capillary Gel Electrophoresis Technique to Genotype CRISPR/Cas9-mediated Knockout Mutants in a High-throughput Format
Published on: April 8, 2017
A deterministic genotyping workflow reduces waste of transgenic individuals by two-thirds
Frederic Strobl1, Ernst H K Stelzer2
1Physical Biology/Physikalische Biologie (IZN, FB 15), Buchmann Institute for Molecular Life Sciences (BMLS), Cluster of Excellence Frankfurt - Macromolecular Complexes (CEF - MC), Goethe-Universität Frankfurt Am Main (Campus Riedberg), Max-von-Laue-Straße 15, 60438, Frankfurt am Main, Germany. frederic.strobl@physikalischebiologie.de.
Abstract:
We present a deterministic workflow for genotyping single and double transgenic individuals directly upon nascence that prevents overproduction and reduces wasted animals by two-thirds. In our vector concepts, transgenes are accompanied by two of four clearly distinguishable transformation markers that are embedded in interweaved, but incompatible Lox site pairs. Following Cre-mediated recombination, the genotypes of single and double transgenic individuals were successfully identified by specific marker combinations in 461 scorings.

