Single DNA molecule denaturation using laser-induced heating

Hung, Min-Sheng; Kurosawa, Osamu; Washizu, Masao

doi:10.1016/j.mcp.2012.03.008

Cited by 17 publications

(15 citation statements)

References 10 publications

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“…Of specific interest in this work is the absorption band near 1440 nm corresponding to the (n 1 n 2 n 3 ) ¼ (1,0,1) combination state for one quantum of both symmetric and asymmetric OH stretching excitation. These IR heating techniques can alleviate many of the limitations associated with the objective-stage heating systems, specifically by providing a broader range of available temperatures and vastly superior spatiotemporal control of temperature (10,12,18,20). Though indirect IR heating techniques have been well described (17), none of the direct techniques have been quantitatively characterized, which represents the first major aim of this work.…”

Section: Introductionmentioning

confidence: 99%

Pulsed IR Heating Studies of Single-Molecule DNA Duplex Dissociation Kinetics and Thermodynamics

Holmstrom

Dupuis

Nesbitt

2014

Biophysical Journal

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Single-molecule fluorescence spectroscopy is a powerful technique that makes it possible to observe the conformational dynamics associated with biomolecular processes. The addition of precise temperature control to these experiments can yield valuable thermodynamic information about equilibrium and kinetic rate constants. To accomplish this, we have developed a microscopy technique based on infrared laser overtone/combination band absorption to heat small (≈10(-11) liter) volumes of water. Detailed experimental characterization of this technique reveals three major advantages over conventional stage heating methods: 1), a larger range of steady-state temperatures (20-100°C); 2), substantially superior spatial (≤20 μm) control; and 3), substantially superior temporal (≈1 ms) control. The flexibility and breadth of this spatial and temporally resolved laser-heating approach is demonstrated in single-molecule fluorescence assays designed to probe the dissociation of a 21 bp DNA duplex. These studies are used to support a kinetic model based on nucleic acid end fraying that describes dissociation for both short (<10 bp) and long (>10 bp) DNA duplexes. These measurements have been extended to explore temperature-dependent kinetics for the 21 bp construct, which permit determination of single-molecule activation enthalpies and entropies for DNA duplex dissociation.

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Section: Introductionmentioning

confidence: 99%

Pulsed IR Heating Studies of Single-Molecule DNA Duplex Dissociation Kinetics and Thermodynamics

Holmstrom

Dupuis

Nesbitt

2014

Biophysical Journal

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“…Subsequently, a laser was used to induce cell lysates and denature DNA. By using an infrared (IR) laser to irradiate DNA, DNA denaturation can be achieved [16][17][18][19][20] and applied to targetsequence detection. 19 Because lasers enable the rapid heating of substances, they can be employed to induce microdroplet PCR [21][22][23] and to investigate PCR within microchannels.…”

Section: Introductionmentioning

confidence: 99%

Laser-induced heating integrated with a microfluidic platform for real-time DNA replication and detection

Hung

Ho²,

Chen

2016

J. Biomed. Opt

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This study developed a microfluidic platform for replicating and detecting DNA in real time by integrating a laser and a microfluidic device composed of polydimethylsiloxane. The design of the microchannels consisted of a laser-heating area and a detection area. An infrared laser was used as the heating source for DNA replication, and the laser power was adjusted to heat the solutions directly. In addition, strong biotin–avidin binding was used to capture and detect the replicated products. The biotin on one end was bound to avidin and anchored to the surface of the microchannels, whereas the biotin on the other end was bound to the quantum dots (Qdots). The results showed that the fluorescent intensity of the Qdots bound to the replicated products in the detection area increased with the number of thermal cycles created by the laser. When the number of thermal cycles was ≥10, the fluorescent intensity of the Qdots was directly detectable on the surface of the microchannels. The proposed method is more sensitive than detection methods entailing gel electrophoresis.

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“…[42] Nukleinsäure-funktionalisierte mesoporçse NPs weisen attraktive Merkmale für die getriggerte Wirkstofffreisetzung auf:1 )V oluminçse DNA-Nanostrukturen, die durch überbrückte Doppelstränge gebildet werden, polymerisierte DNA-Ketten (mittels Hybridisierungskettenreaktion oder polymeraseinduzierter Replikation), der Aufbau von zweioder dreidimensionalen DNA-Strukturen oder die Bildung von Aptamer-Ligand-Komplexen liefern wirksame Nanostrukturen zum Verschließen der Poren. 2) Verschiedene Tr igger,d arunter Strangverdrängung, [7] pH-Wert, [9] Liganden [10] oder thermische Stimuli, [43] Die Entriegelung mesoporçser SiO 2 -NPs durch Rekonfiguration von DNA-Verschlüssen über die Bildung eines Aptamer-Ligand-Komplexes und durch den anschließenden biokatalytischen, Exonuklease-stimulierten Abbau des gebildeten Aptamer-Komplexes wurde für die kontrollierte Freisetzung des Tu mortherapeutikums CPT in Brustkrebszellen vollzogen (Abbildung 3A). [45] Bei diesem System wurde die Tatsache genutzt, dass in Krebszellen eine Überproduktion von ATPe rfolgt und dass EndoGI, ein Nicking-Enzym vom Exonuklease-Typ,i nK rebszellen vorhanden ist.…”

Section: Methodsunclassified

Stimuliresponsive DNA‐funktionalisierte Nano‐ und Mikrocontainer zur schaltbaren und kontrollierten Freisetzung

Lü

Willner

2015

Angewandte Chemie

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Nanotechnology enables the design of materials with outstanding performance. A key element of nanotechnology is the ability to manipulate and control matter on the nanoscale to achieve a certain desired set of specific properties. Here, we discuss recent insight into the formation mechanisms of inorganic nanoparticles during precipitation reactions. We focus on calcium carbonate, and describe the various transient stages potentially occurring on the way from the dissolved constituent ions to finally stable macrocrystals—including solute ion clusters, dense liquid phases, amorphous intermediates, and nanoparticles. The role of polymers in nucleating, templating, stabilizing, and/or preventing these structures is outlined. As a specific example for applied nanotechnology, the properties of cement are shown to be determined by the formation and interlocking of calcium‐silicate‐hydrate nanoplatelets. The aggregation of these platelets into mesoscale architectures can be controlled with polymers.

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Single DNA molecule denaturation using laser-induced heating

Cited by 17 publications

References 10 publications

Pulsed IR Heating Studies of Single-Molecule DNA Duplex Dissociation Kinetics and Thermodynamics

Pulsed IR Heating Studies of Single-Molecule DNA Duplex Dissociation Kinetics and Thermodynamics

Laser-induced heating integrated with a microfluidic platform for real-time DNA replication and detection

Stimuliresponsive DNA‐funktionalisierte Nano‐ und Mikrocontainer zur schaltbaren und kontrollierten Freisetzung

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