A graphene-based platform for fluorescent detection of SNPs

Area of Science:

• Biochemistry
• Analytical Chemistry
• Materials Science

Background:

• Single nucleotide polymorphisms (SNPs) are crucial genetic markers.
• Accurate and sensitive SNP detection methods are essential for diagnostics and research.
• Graphene oxide (GO) possesses unique optical and adsorption properties.

Purpose of the Study:

• To develop a novel, rapid, and sensitive fluorescent assay for single nucleotide polymorphism (SNP) detection.
• To utilize graphene oxide (GO) for SNP discrimination based on differential fluorescence quenching.

Main Methods:

• Development of a single-base extension reaction coupled with a graphene oxide (GO) based fluorescence assay.
• Exploitation of the differential absorption capacities of fluorescein-labeled dGTP (dGTP-Fl) and double-stranded DNA (dsDNA) by GO.
• Discrimination between wild-type and mutant targets based on fluorescence recovery or quenching.

Main Results:

• The assay demonstrated high sensitivity with a detection limit of 3 nM for mutant targets.
• A linear detection range was established from 3 nM to 50 nM for mutant targets.
• The method successfully detected as low as 10% mutant targets in the presence of a 9-fold excess of wild-type targets.

Conclusions:

• The developed GO-based fluorescent assay provides a simple, fast, and sensitive platform for SNP detection.
• This method shows significant potential for genetic analysis and molecular diagnostics.
• The assay's ability to detect low-frequency mutations is advantageous for various applications.