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We demonstrate a simple and rapid method for SNP typing, allele frequency determination, and trace mutant analysis that works with even an inexpensive detection system. This method is based on microchip electrophoresis of tagged probes incorporated with one-colored ddNTP (METPOC). The assay uses dye terminator incorporation into a pair of probes of different lengths specific to wild- and mutant-type targets, respectively. They are hybridized to the targets prior to ddNTP-Cy-5 incorporation, which occurs only for a matched probe-target duplex. Because the extension reactions for the two probes are carried out simultaneously in one tube and the products from both probes are analyzed in one channel by one-color fluorescence detection, an accurate comparative analysis of SNPs is possible. SNP typing as well as allele frequency determination in the range above 0.1% can easily be carried out using a commercial microchip electrophoresis system in a few minutes.

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SynopsisIt has been observed that the viscosity of polyacrylamide (PAM) solution decreased with an increase in high-speed stirring time. The viscosities of PAM solutions decrease initially and then remain constant as the stirring time is increased. The final values depend on the stirring speed. This decrease was assumed to be due to a change in the dissolved state and/or the degradation of PAM molecules. It was discovered through experimentation that the molecular weight decreased, implying that the PAM molecules were degraded.

SynopsisIt has been observed that the viscosity of polyacrylamide (PAM) solution decreased with an increase in high-speed stirring time. The viscosities of PAM solutions decrease initially and then remain constant as the stirring time is increased. The final values depend on the stirring speed. This decrease was assumed to be due to a change in the dissolved state and/or the degradation of PAM molecules. It was discovered through experimentation that the molecular weight decreased, implying that the PAM molecules were degraded.