DNA with all cytosines, thymines, or all pyrimidines of one strand substituted by fluorescently labeled analogs shows diminished solubility in aqueous media and a strong tendency to aggregation that hampers enzymatic downstream processing. In this study, immobilization of fully fluorescently labeled DNA on microarrays was shown to resolve the named problems and to enable successive DNA degradation by exonuclease III. Fluorescence correlation spectroscopy and single-molecule counting for monitoring the course of DNA hydrolysis in real time revealed the virtually processive degradation of labeled DNA that occurred at an average rate of approximately 4 nt/s.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1016/j.ab.2009.12.004 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Rapid and Visual Detection of Urinary Pathogens by Employing Bifunctional Deoxyribonucleic Acid Sensors and Aggregation of Gold Nanoparticles.
Anal Chem
December 2024
NMPA Key Laboratory for Research and Evaluation of Drug Metabolism & Guangdong Provincial Key Laboratory of New Drug Screening & Guangdong-Hongkong-Macao Joint Laboratory for New Drug Screening, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou 510515, China.
A simple, rapid, and visual approach is developed to perform diagnosis of urinary tract infection (UTI) and antimicrobial susceptibility testing (AST) by employing smart bifunctional DNA (bfDNA) sensors, exonuclease III, concatermers of CuO nanoparticles (CuONPs), and gold NPs (AuNPs) aggregation [AuNPs agglutination (AA)], namely, the bfDEC-AA method. The bfDNA sensors serve as probes for identifying 16S rRNA genes of bacterium or 18S rRNA of fungus and as mediators connecting the concatermers of CuONPs. The AA as a signal source is triggered by Cu(I)-catalyzed azide-alkyne cycloaddition click chemistry.
View Article and Find Full Text PDFPhoto-Cross-linked DNA Structures Greatly Improves Their Serum Nuclease Resistances and Gene Knock-In Efficiencies.
Small Methods
December 2024
Hefei National Research Center for Physical Sciences at the Microscale, Department of Polymer Science and Engineering, University of Science and Technology of China, Hefei, 230026, China.
The stabilization and structural integrity of DNA architectures remain significant challenges in their biomedical applications, particularly when inserting functional units into the genome using long single-stranded DNA (lssDNA). To address these challenges, a site-specific photo-cross-linking method is employed. Single-stranded oligonucleotides, containing one or two photosensitive cyanovinylcarbazole nucleoside (K) molecules, are precisely incorporated and cross-linked at the specific sites of ssDNA through base-pairing, followed by rapid UV irradiation at 365 nm.
View Article and Find Full Text PDFRolling Circle Amplification Integrating with Exonuclease-III-Assisted Color Reaction for Sensitive Telomerase Activity Analysis.
ACS Omega
December 2024
Department of Ultrasound, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
Telomerase activation can lead to the escape from cell senescence and immortalization, playing a crucial role in the growth and proliferation of cancer cells. Therefore, the detection of telomerase activity is essential for cancer diagnosis and treatment. Herein, we develop a novel ultrasensitive and visually detectable platform.
View Article and Find Full Text PDFAffordable infectious pathogen detection using a dual-mode biosensor integrating exonuclease III-assisted target recycling amplification with high-throughput 96-well microplate format.
Enzyme Microb Technol
February 2025
Chemical Analysis and Biosensors Research Group, Laboratory of Process Engineering and Environment, Faculty of Sciences and Techniques, Hassan II University of Casablanca, Morocco. Electronic address:
Article Synopsis
- The proposed biosensor provides an affordable platform for detecting infectious pathogens, specifically using Hepatitis B virus (HBV) as a model.
- It utilizes an exonuclease III-assisted amplification strategy in a high-throughput 96-well format, increasing sensitivity for HBV DNA detection through repeated binding cycles.
- With a detection limit of 5.62 fM and successful quantification in human serum, this dual-mode biosensor presents a practical solution for medical diagnostics and public health monitoring.
Rolling circle amplification cooperating crRNA switch for direct and sensitive methicillin-resistant Staphylococcus aureus (MRSA) analysis.
Biotechnol Lett
November 2024
Department of Gastroenterology, People's Hospital Of Chongqing Liang Jiang New Area, No. 199 Renxing Road, Renhe Street, Yubei District, Chongqing, 401147, China.
Evaluating the methicillin resistance of Staphylococcus aureus (S. aureus) is highly important for adapting nursing strategies. Nevertheless, the identification of methicillin-resistant S.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!