Digital microfluidics multi-dynamic range parallel biochemical assays

A method for assaying analytes on a microfluidic device, the method comprising:

• a. loading a sample comprising an analyte to be assayed onto a microfluidic device;
• b. performing a short-incubation reaction protocol to measure a relatively high concentration of the analyte, the short-incubation reaction protocol comprising merging a first droplet of the sample with one or more reaction reagents, wherein the short-incubation reaction protocol is optimized to measure a relatively high concentration of the analyte, and wherein a first relatively high concentration range negative-slope standard curve is used for quantifying the analyte;
• c. performing a long incubation reaction protocol to measure a relatively low concentration of the analyte, the long incubation reaction protocol comprising merging a second droplet of the sample with the one or more reaction reagents, wherein the long incubation reaction protocol is optimized to measure a relatively low concentration of the analyte, and wherein a second relatively low concentration range negative-slope standard curve is used for quantifying the analyte;
• d. optionally, performing an intermediate incubation reaction protocol to measure an intermediate concentration of the analyte, the intermediate incubation reaction protocol comprising merging a third droplet of the sample with one or more reaction reagents, wherein the intermediate reaction protocol is optimized to measure an intermediate concentration of the analyte, and wherein a third intermediate concentration range negative-slope standard curve is used for quantifying the analyte;
• and
• e. calculating a concentration of the analyte using the high concentration range negative-slope standard curve, the low concentration range negative-slope standard curve, and/or optionally the intermediate negative-slope standard curve.