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UV melting experiments

The melting of DNA can be easily monitored by a hyperchromic shift resulting from the unstacking of base bases and dissociation of the duplex DNA. This quick and simple method can also be applied to quadruplex DNA utilizing the same principles. Measuring the spectral absorbance at 295 nm, a maximum for [Pg.25]


As shown by a UV melting experiment, successful triplex formation can be achieved when this inverted triad is accommodated in T AT triplexes. The two A moieties in the inverted repeat are not likely to be identical, and the issue is yet to be resolved as to whether the two have different glycosidic torsions and so on. These examples involving pseudopyrimidines in the central position have really extended the triplex recognition motifs and allow the construction of additional patterns with pyrimidines involved in nonstandard WC pairs. [Pg.298]

The concept of absorbance units at 260 nm, A bo (aIso known as optical density units. ODU) often causes confusion for students. By definition, one unit is the amount of sample that gives a UV absorbance at X = 250 nm of 1.0 if the sample is dissolved in 1 ml volume and measured in a l-cm pathlength cuvette. Since aborbance is proportional to concentration (Beer s Law) and the volume is 1 ml, the number of Ajsq units of a sample is proportional to the number of moles of strands. Knowledge of the number of A260 units is important for planning UV melting experiments to optimize sensitivity. [Pg.338]

Choice of buffer is an important yet often overlooked aspect of performing reliable UV thermal denaturation studies. Among the most important considerations are the buffer pK, buffer capacity, compatibility with divalent metals and temperature dependence of pKg. Table 1 lists the properties of the most commonly used buffers for UV melting experiments. A list of other non-complexing buffers over the entire pH range has recently been described (64). [Pg.339]

Figure 2 Typical experiment UV melting profile. At a given temperature, the fraction of strands in the duplex state, a, is given by the ratio a/(a + b), where a and b are the respective vertical distances from the upper and lower baselines to the experimental melting curve. (Reproduced with permission from Gore MA (ed.) (2000) Spectrophotometry and Spectrofluorimetry A Practical approach, p. 330. Oxford Oxford University Press.)... Figure 2 Typical experiment UV melting profile. At a given temperature, the fraction of strands in the duplex state, a, is given by the ratio a/(a + b), where a and b are the respective vertical distances from the upper and lower baselines to the experimental melting curve. (Reproduced with permission from Gore MA (ed.) (2000) Spectrophotometry and Spectrofluorimetry A Practical approach, p. 330. Oxford Oxford University Press.)...
In a few experiments the oxidation product (disulphides) was precipitated by addition of ethanol followed by cooling to 0 C. The purity of the product as disulphide was checked by melting point, FT-IR and UV-visible spectra and compared with that of authentic disulphide prepared from the corresponding thiols [17]. [Pg.976]


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