TITLE

Rapid, single-step nucleic acid detection

AUTHOR(S)
Cissell, Kyle A.; Campbell, Sean; Deo, Sapna K.
PUB. DATE
August 2008
SOURCE
Analytical & Bioanalytical Chemistry;Aug2008, Vol. 391 Issue 7, p2577
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
ABSTRACT
A rapid detection method for nucleic acid based on bioluminescence resonance energy transfer (BRET) from the luminescence donor Renilla luciferase to an acceptor quantum dot upon oligonucleotide probe hybridization has been developed. Utilizing a competitive assay, we detected the target nucleic acid by correlating the BRET signal with the amount of target present in the sample. This method allows for the detection of as little as 4 pmol (20 nM) of nucleic acid in a single-step, homogeneous format both in vitro in a buffer matrix as well as in a cellular matrix. Using this method, one may perform nucleic acid detection in as little as 30 min, showing much improvement over time-consuming blotting methods and solid-phase methods which require multiple wash steps to remove unbound probe. This is the first report on the use of quantum dots as a BRET acceptor in the development of a nucleic acid hybridization assay.
ACCESSION #
33372834

 

Related Articles

  • Toward a solid-phase nucleic acid hybridization assay within microfluidic channels using immobilized quantum dots as donors in fluorescence resonance energy transfer. Lu Chen; Algar, W. Russ; Tavares, Anthony J.; Krull, Ulrich J. // Analytical & Bioanalytical Chemistry;Jan2011, Vol. 399 Issue 1, p133 

    The optical properties and surface area of quantum dots (QDs) have made them an attractive platform for the development of nucleic acid biosensors based on fluorescence resonance energy transfer (FRET). Solid-phase assays based on FRET using mixtures of immobilized QD-oligonucleotide conjugates...

  • GPCRs: Caught in a spectroscopic trap. Piehler, Jacob // Nature Chemical Biology;Sep2011, Vol. 7 Issue 9, p578 

    The article focuses on the identification of the role of G protein-coupled receptors (GPCR) dimerization in functional selectivity through a spectroscopic assay. It states that the study of E. Urizar and colleagues showed the use of a reporter assay for conformational changes in the GPCR-G...

  • Methods in molecular medicine. Craig, R.K. // British Medical Journal (Clinical Research Edition);9/12/1987, Vol. 295 Issue 6599, p646 

    Presents the concept of nucleic acid hybridization in diagnosis. Overview of deoxyribonucleic acid structure; Techniques required for direct visual comparison of genes; Techniques of hybridization analysis.

  • Structure of a new nucleic-acid-binding motif in eukaryotic transcriptional elongation factor TFIIS. Qian, Xiuqu; Jeon, ChoonJu // Nature;9/16/1993, Vol. 365 Issue 6443, p277 

    Describes the three-dimensional nuclear magnetic resonance (NMR) structure of a Cys4 nucleic-acid-binding domain from human TFIIS. Occurrence of analogous Cys4 structural motifs in other proteins; Growing recognition of beta-sheet as a motif of nucleic-acid recognition.

  • Nucleic acid evolution and minimization by nonhomologous random recombination. Bittker, Joshua A.; Le, Brian V.; Liu, David R. // Nature Biotechnology;Oct2002, Vol. 20 Issue 10, p1024 

    We have developed a simple method for exploring nucleic acid sequence space by nonhomologous random recombination (NRR) that enables DNA fragments to randomly recombine in a length-controlled manner without the need for sequence homology. We compared the results of using NRR and error-prone PCR...

  • Novel observations of Thiobacterium, a sulfur-storing Gammaproteobacterium producing gelatinous mats. Grünke, Stefanie; Lichtschlag, Anna; Beer, Dirk de; Kuypers, Marcel; Lösekann-Behrens, Tina; Ramette, Alban; Boetius, Antje // ISME Journal: Multidisciplinary Journal of Microbial Ecology;Aug2010, Vol. 4 Issue 8, p1031 

    The genus Thiobacterium includes uncultivated rod-shaped microbes containing several spherical grains of elemental sulfur and forming conspicuous gelatinous mats. Owing to the fragility of mats and cells, their 16S ribosomal RNA genes have not been phylogenetically classified. This study...

  • Interfacial Chemistry and the Design of Solid-Phase Nucleic Acid Hybridization Assays Using Immobilized Quantum Dots as Donors in Fluorescence Resonance Energy Transfer. Algar, W. Russ; Krull, Ulrich J. // Sensors (14248220);2011, Vol. 11 Issue 6, p6214 

    The use of quantum dots (QDs) as donors in fluorescence resonance energy transfer (FRET) offer several advantages for the development of multiplexed solid-phase QD-FRET nucleic acid hybridization assays. Designs for multiplexing have been demonstrated, but important challenges remain in the...

  • A new method for the detection of ATP using a quantum-dot-tagged aptamer. Chen, Zhang; Li, Guang; Zhang, Lan; Jiang, Junfeng; Li, Zhao; Peng, Zhihui; Deng, Le // Analytical & Bioanalytical Chemistry;Nov2008, Vol. 392 Issue 6, p1185 

    Fluorescence resonance energy transfer (FRET) between a quantum dot as donor and an organic fluorophore as acceptor has been widely used for detection of nucleic acids and proteins. In this paper, we developed a new method, characterized by 605-nm quantum dot (605QD) fluorescence intensity...

  • Quantum dots as donors in fluorescence resonance energy transfer for the bioanalysis of nucleic acids, proteins, and other biological molecules. Algar, W. Russ; Krull, Ulrich J. // Analytical & Bioanalytical Chemistry;Jul2008, Vol. 391 Issue 5, p1609 

    Quantum dots (QDs) have a number of unique optical properties that are advantageous in the development of bioanalyses based on fluorescence resonance energy transfer (FRET). Researchers have used QDs as energy donors in FRET schemes for the analysis of nucleic acids, proteins, proteases,...

Share

Read the Article

Courtesy of THE LIBRARY OF VIRGINIA

Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics