Figure 1.

Proposed MeltPro HBB assay. A: Genotyping procedure. After DNA extraction, each sample tested is mixed with premixed PCR reagents in two separate tubes. PCR and the subsequent melting curve analysis were performed using a real-time PCR thermal cycler. Graphic output of genotyping results is produced. B: Principle of the MeltPro HBB assay. Asymmetric PCR is used to generate single-strand target sequences for hybridization using dual-labeled self-quenched probes, which are labeled at one of four different fluorophores at 5′-end and a quencher dye at 3′-end. One probe may detect one or several adjacent mutations, and it may also be possible for certain mutations to be detected by more than one probe. Examples shown are heterozygous (red line), wild-type (dashed green line), and homozygous mutant (dashed blue line) samples.