Surface-enhanced Raman spectroscopy (SERS) was utilized for the quantitative analysis of double-stranded (ds) DNA amplified by a polymerase chain reaction (PCR). 4?, 6-Diamidino-2-phenylindole dihydrochloride (DAPI), which intercalates into ds-DNA but does not form a complex with single-stranded (ss) DNA, was added to a DNA solution after amplification by PCR. When the solution was mixed, including ds-DNA-DAPI complexes and free DAPI with silver colloid sol, only free DAPI was adsorbed on the colloid surface. The dye on the colloid gave very intense SERS signals with excitation at 514.5 nm, whereas DAPI engaging in the intercalation with ds-DNA did not show any SERS signal. The SERS spectrum of DAPI on the colloid showed a strong band at 1610 cm(-1) due to the C?N stretching mode, and a linear relationship was observed between the peak intensity of the C?N stretching band and the concentration of free DAPI. Therefore one can determine the concentration of free DAPI by the SERS measurement. The more ds-DNA there is in the solution, the less free DAPI there is. Thus it is possible to quantitatively analyze the ds-DNA amplified by PCR indirectly by using SERS. The correlation coefficient between the peak intensity of the C?N stretching band and the concentration of ds-DNA amplified by PCR was calculated to be 0.988 for a concentration range from 0.1 to 1.3 mg/ml.
We describe the quantitative analysis of some metabolic gases bymultichannel Raman spectroscopy. Raman spectra were measured forair, acetone, ammonia, carbon dioxide, and mixed gas consisting ofacetone, ammonia, and air. We designed a new elliptic-sphericalintegration type of cell holder to obtain the Raman spectra of gaseswith a high signal-to-noise ratio. Concentrations of acetone, ammonia, and carbon dioxide were determined by the peak intensities ofRaman bands at 2940, 3228, and 1385 cm(-1), respectively. To compensate for the fluctuations of Ramanintensities caused by several factors, such as the fluctuations oflaser power, the peak intensity of a band at 2324 cm(-1) dueto nitrogen gas was used as an internal intensity standard. Thecorrelation coefficient between the corrected Raman intensity at 2940cm(-1) and the concentration of acetone was calculated to be0.984 for a concentration range of 2-12 ppm. The detection limitof acetone gas was found to be 2 ppm.
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