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Figure 1 | Malaria Journal

Figure 1

From: Real-time PCR/MCA assay using fluorescence resonance energy transfer for the genotyping of resistance related DHPS-540 mutations in Plasmodium falciparum

Figure 1

Schematic representation of FRET assay. Step 1. A PCR reaction results in amplification of a 169 bp fragment with an incorporated ROX fluorophore attached to the forward primer. Step 2. After amplification, the FAM-labelled probe is added to the reaction and during initial denaturation hybridizes to the amplicon. Step 3. The two fluorophores are now in close proximity of each other and energy from the excited donor is transferred to the acceptor generating the FRET signal. Step 4. The increase of temperature during melt curve analysis leads, at a specific temperature, to the dissociation of the probe from the amplicon. When the probe is dissociated transfer of energy is lost and therefore no FRET signal can be observed.

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Malaria Journal

ISSN: 1475-2875

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