Solvent Effect to the Uniformity of Surfactant-Free Salmon-DNA Thin Films

Fabrication of surfactant-modified DNA thin films with high uniformity, specifically DNA–CTMA, has been well considered via drop-casting and spin-coating techniques. However, the fabrication of thin films with pure DNA has not been sufficiently studied. We characterize the uniformity of thin films from aqueous salmon DNA solutions mixed with ethanol, methanol, isopropanol, and acetone. Measurements of thickness and macroscopic uniformity are made via a focused-beam ellipsometer. We discuss important parameters for optimum uniformity and note what the effects of solvent modifications are. We find that methanol- and ethanol-added solutions provide optimal fabrication methods, which more consistently produce high degrees of uniformity with film thickness ranging from 20 to 200 nm adjusted by DNA concentration and the physical parameters of spin-coating methods.

Molecular Transformations in DNA under the Influence of UV-radiation

Background: The processes of destruction and crosslinking of macromolecules occur simultaneously under the influence of ultraviolet (UV) radiation in synthetic polymers, dry DNA and their concentrated solutions. Objective: The effect of UV radiation on calf thymus DNA in dilute solutions subjected to UV- irradiation was studied in this work. Method: The calf thymus DNA was studied in dilute solutions using viscometry, absorption spectroscopy and electrophoresis. Results: It was shown, that at a low concentration of DNA in the buffer solution ([DNA] = 85 μg / ml) under the influence of UV radiation, the processes of destruction of macromolecules and an increase in their flexibility predominate, which is accompanied by a gradual decrease in the viscosity of their solution. In addition, due to the low concentration of the solution, intramolecular crosslinking of macromolecules predominates, which also reduces their size and, consequently, the viscosity of the solution. Conclusion: It was concluded, that in dilute DNA solutions, due to the predominance of the processes of intramolecular crosslinking of macromolecules over intermolecular, only constant processes of decreasing the sizes of DNA macromolecules occur. As a result, its solubility remains virtually unchanged during UV irradiation. The described comments are also excellently confirmed by the results of absorption spectroscopy and electrophoresis

Bleaching correction for DNA-measurements in highly diluted solutions using confocal microscopy

Precise and reliable determination of the nucleic acid concentration in biological samples still remains a challenge. This is particularly since the established fluorescene-based methods provide insufficient results, when only minute sample quantitites are available for analysis. Among other effects, photobleaching is the main reason for this. Since large molecules diffuse more slowly than small molecules, they are exposed to more excitation cycles and therefore have a higher probability of permanently losing their fluorescence. Solutions with large molecules hence show a reduced fluorescence. In this paper we present a method to correct this effect and thus allow high-precision sample concentration determination in minute sample quantities (< 2 µl drops with concentrations < 20 pg/µl). For this purpose, we used confocal microscopy with single molecule sensitivity. In the first step, we derived calibration curves from DNA solutions with defined fragment length. We analyzed dilution series over a wide concentration range (1 pg/µl – 1000 pg/µl) and measured their specific diffusion coefficients by employing fluorescence correlation spectroscopy. Using this information, we correct the measured fluorescence intensity of the calibration solutions for photobleaching effects. Subsequently, we evaluated our method by analyzing a series of DNA mixtures of varying composition.

Molecular Spectroscopic Markers of DNA Damage

Every cell in a living organism is constantly exposed to physical and chemical factors which damage the molecular structure of proteins, lipids, and nucleic acids. Cellular DNA lesions are the most dangerous because the genetic information, critical for the identity and function of each eukaryotic cell, is stored in the DNA. In this review, we describe spectroscopic markers of DNA damage, which can be detected by infrared, Raman, surface-enhanced Raman, and tip-enhanced Raman spectroscopies, using data acquired from DNA solutions and mammalian cells. Various physical and chemical DNA damaging factors are taken into consideration, including ionizing and non-ionizing radiation, chemicals, and chemotherapeutic compounds. All major spectral markers of DNA damage are presented in several tables, to give the reader a possibility of fast identification of the spectral signature related to a particular type of DNA damage.