( nt probe not only hybridized...)The Gradient Feature: Use in Optimization of Allelic Discrimination Assays, Rev B

In summary, we have demonstrated that the gradient feature is a powerful tool for the optimization of PCR protocols. Useful technical information such as the optimal T_a for obtaining maximal PCR product yield (Rychlik et al. 1990) or the maximal temperature at which PCR works (Wu et al. 1991) can be generated using the gradient in a single experiment. The gradient not only dramatically accelerates the PCR T_a optimization process but also the optimization of SNP detection protocols. In addition to TaqMan and molecular beacons, the iCycler iQ gradient feature can be used to optimize genotypic assays that involve allele-specific PCR (Whitcombe et al. 1999) and fluorescence resonance energy transfer (FRET) using dual-labeled probes (van den Bergh et al. 2000).

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