Improved signal-to-noise ratio with FRET-based hybridization probes
Molecular Beacons are dual-labeled probes that form a quenched, stem-loop structure in native state and fluoresce upon hybridization to the target nucleotide sequence. Applications include real-time and endpoint PCR, SNP detection, and multiplex amplification.
Prices listed include probe sequence (up to 45 bases in length), reporter, quencher, and HPLC purification. Turnaround time is 5–7 business days for standard molecular beacon probes.
To request molecular beacon probes with <18 or >45 bases, please contact our Technical and Customer Support.
Step 1: During the annealing step, hybridization of molecular beacon probes to target sequences separate the fluorescent dye (D) and quencher (Q), resulting in fluorescence that is detected by the real-time PCR instrument. Probes that do not hybridize to target DNA sequences will reform the hairpin structure and will not fluoresce.
Step 2: The polymerase extends from the primers and begins DNA synthesis.
Step 3: When the polymerase reaches the molecular beacon, the probe is displaced without being degraded. Therefore, molecular beacons can participate in multiplex rounds of annealing.
Step 4: The polymerase continues extension of the primers to complete synthesis of the DNA strand.