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DPO™ Technology is the innovative multiple targets amplification technology that enhances target specificity and minimizes the non-specific amplification commonly occurred in multiplex PCR. DPO™ technology redefines high multiplex PCR by enabling to detect “many targets in a single tube” and is well-suited for high multiplex real-time PCR technology.
The principle of DPO™ technology
DPO™ primer is structurally different from the conventional primer and is composed of two functional priming portions connected by the Poly-dI linker. Length of 5’-end target stabilizing portion is longer than that of 3’-end target determinant portion. Poly-dI linker forms a bubble-like structure at a certain annealing temperature and controls the two-step priming reactions required for polymerase to extend a DPO™primer. With a DPO™ primer, initial binding is through 5’-end portion and then primer extension is controlled through more specific annealing of the 3’-end portion.
Step 1: First priming reaction
The longer 5'- end portion preferentially binds to the template DNA and initiates "stable annealing".
Step 2: Second priming reaction
The short 3'- end portion selectively binds to a target site and determines "target-specific extension”.
Unparalleled specificity by eliminating any false extension
DPO™ primer does not generate any false positive signal by blocking the extension of primer on non-target template, thus provides unparalleled specificity in high multiplex PCR.
A. Although the longer 5’- end portion binds a non-target site, the short potion resists non-specific extension.
B. The short 3’- end portion alone fails to make a priming at an annealing temperature.
DPO™ multiplex PCR assay is the most accurate, rapid, and cost effective method.
Example
Comparison of Multiplex PCR results with DPO™ primer vs. conventional primer
DPO™ primer eliminated non-specific amplification and produced only true target bands.
However, conventional primer did not eliminate nonspecific amplification.
A. Conventional primers
P: Positive marker N: Negative marker / Lane 1~5: Samples
B. DPO™ primers
P: Positive marker N: Negative marker / Lane 1~5: Samples