Figure 1

SNP typing by PrASE. A nested multiplex PCR is performed to amplify all SNP loci in a single reaction. The biotin-labeled inner PCR products are captured by streptavidin-coated magnetic beads facilitating automated reaction clean-ups between all assay steps. Strand-specific alkali elution is then performed before hybridization of allele-specific extension primers which contain unique tag sequences for later microarray detection. The multiplex PrASE reaction is performed with Cy5-labeled dNTPs to facilitate fluorescence detection. The products of the reaction are released with alkali, neutralized, and hybridized to a universal tag microarray containing 48 identical wells before detection.