Fluorescence-Based Detection of <i>KRAS</i> Mutations in Genomic DNA Using Magnetic Bead-Coupled LDR Assay.

ABSTRACT: We previously developed a bead-coupled ligase detection reaction (LDR) assay that enables simple and rapid detection of single-nucleotide variations (SNVs) using synthetic oligonucleotide templates. In the present study, this approach was extended to genomic DNA extracted from colorectal cancer cell lines to evaluate its applicability to clinically relevant samples. Targeting codon 12 of the KRAS gene, PCR-amplified products served as templates for bead-coupled LDR, and fluorescence excitation-emission matrix (EEM) analysis was employed for signal readout. The four fluorophores used in the assay exhibited distinct spectral properties, allowing their signals to be clearly resolved within the EEM profiles. This mapping provided characteristic fluorescence signatures that revealed the

SUBMITTER: Morimoto C 

PROVIDER: S-EPMC12736157 | biostudies-literature | 2025 Dec

REPOSITORIES: biostudies-literature