Homologous Recombination Confirmation in ES Cells

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The Mouse Biology Program offers specialized services for confirming homologous recombination in embryonic stem (ES) cells, a crucial step in creating targeted gene modifications. This confirmation ensures that genetic alterations have been correctly integrated at the desired locus, maintaining the integrity and accuracy of engineered mouse models.

What is Homologous Recombination?

Homologous recombination in ES cells is a gene-targeting technique that introduces specific genetic modifications, such as knockouts, knock-ins, or conditional alleles. This method uses homologous DNA sequences to guide the insertion or replacement of DNA at the target site, allowing for precise genetic changes.

Homologous Recombination Confirmation Process

• PCR Screening:
  Polymerase Chain Reaction (PCR) is used as an initial screen to detect the presence of correctly integrated alleles. Primers designed to bind specifically to the modified region confirm successful recombination at the target site.
• Southern Blot Analysis:
  Southern blotting provides a higher level of confirmation by detecting specific DNA fragments and verifying the presence and structure of targeted alleles. Visualizing the integration pattern helps identify correctly targeted ES cell clones.
• Sequencing of Targeted Loci:
  Sanger sequencing confirms the precise nucleotide changes at the targeted locus, ensuring that the recombination event occurred accurately and without unintended mutations.

Why is Confirmation Important?

• Ensuring Genetic Integrity:
  It verifies that the genetic modification has occurred as designed, preventing off-target effects or unintended genetic changes.
• Increasing Success Rates:
  Confirming successful targeting in ES cells improves the efficiency of generating genetically modified mouse models, saving time and resources.
• Maintaining Research Validity:
  Accurate confirmation supports reliable research outcomes by ensuring that only correctly targeted cells are used in subsequent model development.

How to Request This Service

Please contact our team to request homologous recombination confirmation in ES cells. We will guide sample submission, service timelines, and expected results, ensuring your genetic modification project meets the highest standards.

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• Genotyping Services
• Coding Sequence Validation
• Genetic Analysis Services
• Genomic Insertion Site Identification
• Homologous Recombination Confirmation in ES Cells
• Off-Target Mutagenesis Detection