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MeltPlex® Method

Here you'll see the MeltPlex® multiplex PCR process 

 

1. Hybridization

PCR primers for amplification of DNA or reverse transcribed RNA target serve as target DNA. An amplification step is performed using PCR primers and multiple target-specific MeltPlex® probes labeled with fluorescent dyes.

2. Probe cleavage

If the specific target is present, the 5’ quencher is cleaved from the probe which is now activated (fluorescent).

 

 

 

3. Detection

A second, common, quenching probe can hybridize to the Melting Tag region and generate a melting curve with a TM specific for each target-specific probes.

 

 

 

4. Read-out

The detection of the activated probes is done by melting curve analysis on a Real-Time PCR system. 20+ target-specific MeltPlex® detection probes, labeled by multiple fluorophores and varying in Melting Tag temperature, enable specific detection of amplified product and the corresponding pathogen.