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A Broadly Applicable Assay for Rapidly and Accurately Quantifying DNA Surface Coverage on Diverse Particles
Haixiang Yu1, Xiaowen Xu1, Pingping Liang1
1Department of Chemistry and Biochemistry, Florida International University , 11200 SW Eighth Street, Miami, Florida 33199, United States.
Bioconjugate Chemistry
|February 4, 2017
Summary
A new assay accurately quantifies DNA surface coverage on particles using exonuclease III (Exo III) digestion. This method is broadly applicable across various particle types and DNA conjugation chemistries for improved sensing and material science applications.
Area of Science:
- Bioconjugation Chemistry
- Nanomaterials Science
- Molecular Diagnostics
Background:
- DNA-modified particles are crucial for sensing, material science, and molecular biology.
- Accurate quantification of DNA surface coverage is essential for particle performance.
- Existing quantification methods have limitations in scope and applicability.
Purpose of the Study:
- To develop a broadly applicable assay for quantifying DNA surface coverage on diverse particles.
- To establish a simple and accurate method using exonuclease III (Exo III) digestion.
- To overcome limitations of current DNA quantitation techniques.
Main Methods:
- Utilized a novel exonuclease III (Exo III) digestion assay targeting phosphodiester bonds.
- Employed fluorophore-labeled DNA probes with abasic sites hybridized to complementary DNA (cDNA).
- Quantified DNA surface coverage based on the release of fluorophores upon enzymatic digestion.
Main Results:
- Demonstrated accurate, rapid, and reproducible DNA quantitation on gold nanoparticles.
- Validated the assay's broad applicability across different particle compositions, sizes, and conjugation chemistries.
- Showcased the assay's efficiency through a cyclic hybridization-digestion process.
Conclusions:
- The developed Exo III digestion assay provides a versatile and accurate tool for DNA surface coverage quantitation.
- This method significantly advances the characterization of DNA-modified particles for various applications.
- Offers a universally applicable solution for researchers working with DNA-modified materials.
