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Validation, QC & Genotyping

Sanger / NGS Validation of Junctions

Sequencing tools for verifying the accuracy of DNA junctions at targeted integration sites. These assays confirm that homologous recombination or HDR occurred precisely without insertions, deletions, or rearrangements.

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Overview

Sanger sequencing and next-generation sequencing (NGS) are essential tools for verifying the accuracy of DNA junctions at targeted integration sites. These assays confirm that homologous recombination or HDR occurred precisely without insertions, deletions, or rearrangements.

Frequently Asked Questions

When is NGS preferred over Sanger sequencing?

NGS offers deep coverage for detecting minor allelic variants and is valuable for humanized constructs and large-fragment integrations where Sanger reads are insufficient.

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Related Terms

Southern Blot / Copy Number Confirmation

A classical DNA analysis method that remains the gold standard for verifying correct targeting, integration patterns, and copy number of inserted constructs in engineered mouse models.

Off-Target Assessment / Allele Integrity

Procedures to determine whether unintended genomic changes have occurred during editing, while also verifying that the targeted locus remains structurally sound and functionally unaffected.

Homology-Directed Repair (HDR)

A high-fidelity DNA repair mechanism that uses a homologous DNA sequence as a template to accurately fix double-strand breaks. Unlike NHEJ, HDR ensures precise integration or correction of genetic material.

More in Validation, QC & Genotyping

Genotyping PCR / qPCRSouthern Blot / Copy Number ConfirmationOff-Target Assessment / Allele IntegrityGermline Confirmation / Coat Color MarkersBreeding Scheme (Hetero × Hetero, etc.)
Safe-Harbor Targeted TransgenesisView All TermsSelection Markers (NeoR, PuroR)

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