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Updated: Apr 18, 2026
Identification of Homologous Recombination Events in Mouse Embryonic Stem Cells Using Southern Blotting and Polymerase Chain Reaction
Published on: November 20, 2018
Multiplex ligation-dependent probe amplification for identification of correctly targeted murine embryonic stem cell
Ewelina Elert-Dobkowska1, J Christopher Hennings2, Christian A Hübner2
1Department of Clinical Chemistry and Laboratory Diagnostics, Jena University Hospital, 07747 Jena, Germany; Department of Genetics, Institute of Psychiatry and Neurology, 02-957 Warsaw, Poland.
Abstract:
Following locus-specific genome editing of mouse embryonic stem cells (ESCs), the identification of correctly targeted clones remains a challenge. We applied multiplex ligation-dependent probe amplification (MLPA) to screen for homologous recombination-based genomic integration of a knockout construct in which part of a gene is deleted. All candidate ESCs thereby identified were subsequently validated by conventional methods. Thus, MLPA represents a highly reliable as well as cost- and time-efficient alternative to currently applied methods such as Southern blotting and polymerase chain reaction (PCR)-based approaches. It is also applicable to knockin recombination strategies and compatible with the CRISPR/Cas9 system and other genome editing strategies.
