Quantitative DNA hybridization in solution using magnetic/luminescent core–shell nanoparticles

Nanoscale magnetic/luminescent core–shell particles were used for DNA quantification in a hybridization-in-solution approach. We demonstrated a rapid, simple, and non-polymerase chain reaction-based DNA hybridization-in-solution assay for quantifying bacteria capable of biodegrading methyl tertiary-...

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Bibliographic Details
Published in:Analytical biochemistry 2007-11, Vol.370 (2), p.186-194
Main Authors: Son, Ahjeong, Dosev, Dosi, Nichkova, Mikaela, Ma, Zhiya, Kennedy, Ian M., Scow, Kate M., Hristova, Krassimira R.
Format: Article
Language:English
Subjects:
Base Pair Mismatch
Base Sequence
DNA
DNA - analysis
DNA - genetics
Hybridization-in-solution
Kinetics
Lanthanide oxide
Luminescence
Magnetic particles
Magnetics
Methyl tertiary-butyl ether
Models, Molecular
Molecular Sequence Data
Nanoparticles
Nucleic Acid Hybridization
Solutions
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Summary:Nanoscale magnetic/luminescent core–shell particles were used for DNA quantification in a hybridization-in-solution approach. We demonstrated a rapid, simple, and non-polymerase chain reaction-based DNA hybridization-in-solution assay for quantifying bacteria capable of biodegrading methyl tertiary-butyl ether. Fe 3O 4/Eu:Gd 2O 3 core–shell nanoparticles synthesized by spray pyrolysis were biofunctionalized with NeutrAvidin. Following immobilization of a biotinylated probe DNA on the particles’ surfaces via passive adsorption, target DNA labeled with fluorescein isothiocyanate was hybridized with probe DNA. The hybridized DNA complex was separated from solution with a magnet, while nonhybridized DNA remained in solution. The normalized fluorescence (fluorescein isothiocyanate/nanoparticles) measured with a spectrofluorometer indicated a linear quantification ( R 2 = 0.98) of the target bacterial 16 S rDNA. The rate of hybridization increased concurrently with the target DNA concentration. In addition, this approach differentiated between the signal outputs from perfectly complementary target and two-base mismatched target DNA in a range of concentrations, showing the specificity of the assay and the possibility for environmental applications.
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2007.08.001