Methods for digital readout quantification of nucleic acids

A method for conducting a quantitative end-point nucleic acid detection via LAMP, said method comprising:

• providing a target sequence;
• providing a reference sequence comprising a tag region, wherein the reference sequence is identical to the target sequence but the tag region is in reverse orientation with respect to a corresponding region in the target sequence;
• providing a first primer which binds with the target sequence;
• providing a second primer which binds with the reference sequence;
• and
• conducting competitive reactions between the reference sequence and the target sequence.

A method for producing a digital readout of nucleic acid detection, said method comprising:

• providing a sample target sequence;
• providing a reference sequence comprising a plurality of sequences identical to said target sequence and a tag region that is in reverse orientation with respect to a corresponding region in the target sequence;
• providing a first loop primer that binds with the sample target, said first loop primer having a first fluorescent molecule attached to its 5′ end;
• providing a second loop primer that binds with the reference target, said second loop primer having a second fluorescent molecule attached to its 5′ end;
• conducting competitive reactions using the sample target sequence and the reference sequence in a set of wells, wherein the concentration of the sample target sequence is equal across the wells and the the concentration of the reference sequence varies from across the wells;
• and
• analyzing the color and intensity of the fluorescence of the wells.