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Dna absorbance spectrum11/9/2023 ![]() Here, it is assumed that there is a state of equilibrium between the 2 different single-stranded (A 1, A 2) and double-stranded (A1A2) states, and that the concentrations of A 1 and A 2 are the same. Thermal stability of nucleic acid is an important factor that controls the structure, hybridization, and functions of nucleic acids therefore, thermal stability analysis of nucleic acid (Tm analysis) is indispensable in these fields.īelow is a simple description of nucleic acid thermal stability analysis (Tm analysis). In addition, nucleic acids (functional nucleic acids) which possess functionality associated with siRNA, antisense nucleic acids, antigene nucleic acids, ribosomes, etc., are applied in various fields, including drug discovery research, drugs, and diagnostics. Hybridization of nucleic acids is utilized in PCR, DNA microarray, southern hybridization, northern hybridization, and in situ hybridization, etc. The phenomenon is referred to as hybridization. In contrast to the melting process, lowering of the temperature will cause a single-stranded nucleic acid to adopt a double-stranded structure through the reinstatement of the hydrogen bonds between the complementary base pairs. Tm is an index of the thermal stability of a nucleic acid, and is dependent on such conditions as the base sequence, base number, nucleic acid concentration, solvent conditions (salt composition, organic solvent composition, pH), mismatch (non-complementary base pairs), nucleic acid analog (artificial nucleic acid) structure, etc. At the melting temperature (Tm), the ratio of the double strands to the single strands becomes equal. In addition to UV absorbance measurement, melting curves are also obtained in other types of measurement, such as circular dichroism spectroscopy and thermal measurement. The data curve acquired when measuring UV absorbance with respect to temperature rise is called a UV absorbance melting curve. Nucleic acids display an ultraviolet absorption peak at 260 nm, and their absorbance at 260 nm increases when melting occurs. This phenomenon is referred to as "melting". When the temperature is raised, the hydrogen bond is broken, the double strand dissociates, and a single-stranded structure is formed. The double strand comprises a sense strand and an antisense strand made up of the complementary bases, G (guanine) and C (cytosine), and A (adenine) and T (thymine), respectively, and these complementary base pairs are joined by a hydrogen bond. DNA typically has a double-stranded helix structure.
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