• The present patent technology aims to disclose a method for quantifying nucleic acids using an Aptamer-based qPCR (Apt-qPCR) probe in real-time PCR.
• This method appears to involve utilizing a light-up dye-aptamer system, where the fluorescence increases significantly when the dye binds to its specific aptamer.

The method for direct quantification of nucleic acids in real-time qPCR, comprises:

• using a simple and shorter Aptamer-based qPCR (Apt-qPCR) probe for quantification in real-time PCR wherein the probe uses a light-up dye-aptamer system in which the dye shows negligible fluorescence in the free state and its fluorescence increases manifold when it binds to its specific aptamer;
• placing the aptamer 5’ upstream of one of the primers wherein the primer initially in pre-annealed form shows fluorescence as aptamer is free and single-stranded & can bind to dye;
• performing annealing and extension step to make the aptamer double-stranded & thereby loose its 3D structure to form a double helix wherein the double helix is not specific for the dye and do not bind, therefore reducing fluorescence of the solution corresponding to each cycle of the PCR reaction.