MultiCode^®-RTx can support single tube multiplex reactions and utilize controls to monitor the reaction. Primer pairs are designed to include a fluorescent reporter-labeled primer with an isoC on the 5' end and an unlabeled primer. Generally, target and control primers are labeled with FAM and HEX respectively.

MultiCode isoC:isoG bases hydrogen bond only with each other and are site-specifically incorporated during amplification. Superior molecular recognition between isobases combined with an elegant two primer system enables detection of almost any nucleic acid target.

Features:

• Probe-free, real-time PCR technology provides reliable results with rapid turn around time.
• Universal cycling conditions and standardized reagents streamline assay workflow and simplify implementation.
• Compatible with most real-time PCR instrumentation.
• Melt curve analysis for confirmation of target amplification and detection of multiple analytes.
• Controls for monitoring extraction and amplification.

	Step 1: Primer Annealing The reporter-labeled forward primer containing a single isoC on the 5' end and unlabeled reverse primer hybridize to the target nucleic acid.
	Step 2: Primer Extension During the amplification process, the labeled primer is incorporated into the newly synthesized strand and serves as a template for the reverse primer in the next cycle.
	Step 3: Site Specifc Incorporation Synthesis of the opposite strand terminates with the incorporation of an isoG with a covalently attached quencher molecule. The resulting proximity of the quencher to the reporter produces a decrease in fluorescence. The fluorescence decrease is directly proportional to the amount of amplicon.
	Step 4: Thermal Melt Following the completion of amplification, a thermal melt is performed and fluorescence is restored after the strands separate.