Multicolor instrumentation for direct fluorescent detection of nucleic acids in a microchip format

1998 ◽  
Author(s):  
Valery L. Bogdanov ◽  
Yu-Hui Rogers ◽  
Guang Lan ◽  
Michael Boyce-Jacino
Keyword(s):  
Nucleic Acids ◽  
Fluorescent Detection

scholarly journals An LED-Driven AuNPs-PDMS Microfluidic Chip and Integrated Device for the Detection of Digital Loop-Mediated Isothermal DNA Amplification

Micromachines ◽  
2020 ◽  
Vol 11 (2) ◽  
pp. 177 ◽  
Author(s):  
Zengming Zhang ◽  
Shuhao Zhao ◽  
Fei Hu ◽  
Guangpu Yang ◽  
Juan Li ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Microfluidic Chip ◽  
Cross Flow ◽  
Flow Channel ◽  
Isothermal Amplification ◽  
Fluorescent Detection ◽  
Lamp Method ◽  
Biological Studies ◽  
Micro Cavity ◽  
Integrated Device

The sensitive quantification of low-abundance nucleic acids holds importance for a range of clinical applications and biological studies. In this study, we describe a facile microfluidic chip for absolute DNA quantifications based on the digital loop-mediated isothermal amplification (digital LAMP) method. This microfluidic chip integrates a cross-flow channel for droplet generation with a micro-cavity for droplet tiling. DNA templates in the LAMP reagent were divided into ~20,000 water-in-oil droplets at the cross-flow channel. The droplets were then tiled in the micro-cavity for isothermal amplification and fluorescent detection. Different from the existing polydimethylsiloxane (PDMS) microfluidic chips, this study incorporates gold nanoparticles (AuNPs) into PDMS substrate through silica coating and dodecanol modification. The digital LAMP chip prepared by AuNPs-PDMS combines the benefits of the microstructure manufacturing performance of PDMS with the light-to-heat conversion advantages of AuNPs. Upon illumination with a near infrared (NIR) LED, the droplets were stably and efficiently heated by the AuNPs in PDMS. We further introduce an integrated device with a NIR heating unit and a fluorescent detection unit. The system could detect HBV (hepatitis B virus)-DNA at a concentration of 1 × 101 to 1 × 104 copies/μL. The LED-driven digital LAMP chip and the integrated device; therefore, demonstrate high accuracy and excellent performance for the absolute quantification of low-abundance nucleic acids, showing the advantages of integration, miniaturization, cost, and power consumption.

scholarly journals Interaction of cyanine dyes with nucleic acids. Meso-substituted trimethincyanines for fluorescent detection of nucleic acids

2002 ◽  
Vol 18 (3) ◽  
pp. 243-252 ◽  
Author(s):  
S. S. Lukashov ◽  
M. Yu. Losytskyy ◽  
Yu. P. Kovtun ◽  
S. M. Yarmoluk
Keyword(s):  
Nucleic Acids ◽  
Cyanine Dyes ◽  
Fluorescent Detection

scholarly journals Symmetric exponential amplification reaction-based DNA nanomachine for the fluorescent detection of nucleic acids

RSC Advances ◽  
2019 ◽  
Vol 9 (70) ◽  
pp. 41305-41310
Author(s):  
Qi Yan ◽  
Qiuyue Duan ◽  
Yuqi Huang ◽  
Jing Guo ◽  
Liang Zhong ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Fluorescent Detection ◽  
Label Free ◽  
Background Signal ◽  
Low Background ◽  
Amplification Reaction ◽  
Dna Nanomachine

This label-free and ultra-low background signal DNA nanomachine was based on P-HP mediated S-EXPAR and the G-triplex/ThT complex.

scholarly journals Fluorescent detection upon droplet amplification of nucleic acids

2019 ◽  
Vol 1410 ◽  
pp. 012079
Author(s):  
A Tupik ◽  
A Bulyanitsa ◽  
G Rudnitskaya ◽  
Y Posmitnaya ◽  
D Varlamov ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Fluorescent Detection

Ultrasensitive fluorescent detection of nucleic acids based on label-free enzymatic-assisted cascade signal amplification

2018 ◽  
Vol 1039 ◽  
pp. 91-97 ◽  
Author(s):  
Yingfeng Qin ◽  
Suqi Liao ◽  
Yufeng Huang ◽  
Jingjin Zhao ◽  
Shulin Zhao
Keyword(s):  
Nucleic Acids ◽  
Signal Amplification ◽  
Fluorescent Detection ◽  
Label Free

Ultrasensitive detection of nucleic acids by template enhanced hybridization followed by rolling circle amplification and catalytic hairpin assembly

2015 ◽  
Vol 51 (12) ◽  
pp. 2392-2395 ◽  
Author(s):  
Weiling Song ◽  
Qiao Zhang ◽  
Wenbo Sun
Keyword(s):  
Nucleic Acids ◽  
Rolling Circle Amplification ◽  
Fluorescent Detection ◽  
Amplification Efficiency ◽  
Ultrasensitive Detection ◽  
Rolling Circle ◽  
Catalytic Hairpin Assembly

An ultrasensitive protocol for fluorescent detection of DNA is designed by combining the template enhanced hybridization process (TEHP) with Rolling Circle Amplification (RCA) and Catalytic Hairpin Assembly (CHA), showing a remarkable amplification efficiency.

scholarly journals Quantitative Capillary Electrophoresis for Analysis of Extracellular Vesicles (EVqCE)

Separations ◽  
2021 ◽  
Vol 8 (8) ◽  
pp. 110
Author(s):  
Yuchu Dou ◽  
Lixuan Ren ◽  
Prabir Kumar Kulabhusan ◽  
Emil A. Zaripov ◽  
Maxim V. Berezovski
Keyword(s):  
Capillary Electrophoresis ◽  
Nucleic Acids ◽  
Cancer Cells ◽  
Extracellular Vesicles ◽  
Dynamic Range ◽  
Fluorescent Detection ◽  
Average Mass ◽  
Promising Tool ◽  
Aggressive Cancer ◽  
Capillary Zone

Extracellular Vesicles (EVs) gained significant interest within the last decade as a new source of biomarkers for the early detection of diseases and a promising tool for therapeutic applications. In this work, we present Extracellular Vesicles Quantitative Capillary Electrophoresis (EVqCE) to measure an average mass of RNA in EVs, determine EV concentrations and the degree of EV degradation after sample handling. We used EVqCE to analyze EVs isolated from conditioned media of three cancer cell lines. EVqCE employs capillary zone electrophoresis with laser-induced fluorescent detection to separate intact EVs from free nucleic acids. After lysis of EVs with a detergent, the encapsulated nucleic acids are released. Therefore, the initial concentration of intact EVs is calculated based on a nucleic acid peak gain. EVqCE works in a dynamic range of EV concentrations from 108 to 1010 particles/mL. The quantification process can be completed in less than one hour and requires minimum optimization. Furthermore, the average mass of RNA was found to be in the range of 200–400 ag per particle, noting that more aggressive cancer cells have less RNA in EVs (200 ag per particle) than non-aggressive cancer cells (350 ag per particle). EVqCE works well for the degradation analysis of EVs. Sonication for 10 min at 40 kHz caused 85% degradation of EVs, 10 freeze-thaw cycles (from −80 °C to 22 °C) produced 40%, 14-day storage at 4 °C made 32%, and vortexing for 5 min caused 5% degradation. Presently, EVqCE cannot separate and distinguish individual EV populations (exosomes, microvesicles, apoptotic bodies) from each other. Still, it is tolerant to the presence of non-EV particles, protein-lipid complexes, and protein aggregates.

Electron Microscopy of Nucleic Acids

1974 ◽  
Vol 32 ◽  
pp. 302-303
Author(s):  
Norman Davidson
Keyword(s):  
Electron Microscopy ◽  
Nucleic Acids ◽  
Basic Protein ◽  
Molecular Biologist ◽  
Topological Properties ◽  
Protein Film ◽  
Polynucleotide Chain

The basic protein film technique for mounting nucleic acids for electron microscopy has proven to be a general and powerful tool for the working molecular biologist in characterizing different nucleic acids. It i s possible to measure molecular lengths of duplex and single-stranded DNAs and RNAs. In particular, it is thus possible to as certain whether or not the nucleic acids extracted from a particular source are or are not homogeneous in length. The topological properties of the polynucleotide chain (linear or circular, relaxed or supercoiled circles, interlocked circles, etc. ) can also be as certained.

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