A cost effective MGB free stem-loop qPCR strategy for high precision microRNA profiling

Numerous techniques are already accessible to measure the expression levels of particular microRNAs in biological specimens. One of the most popular methods and, according to experts in this field, the most accurate method available is the Stem Loop method. In this method, the cDNA of each microRNA is first synthesized explicitly with an adapter’s help. Finally, the measurement uses general reverse primers, specific forward primers, and a specific probe. One of the problems with this method is that the probe binding site is such that in order to have the appropriate melting temperature (Tm), in most cases, it is necessary to bind the MGB molecule to the probe. This protocol requires much expense and significantly increases the cost of the test. In our innovative method, the adapter and the experiment are designed so that to synthesize a suitable probe, there is no need to add the MGB molecule to it, and the probe alone will have the appropriate Tm.