To begin, design DNAzyme-based nano machines, or DNM, with a 10 to 23 core, using free software. Assemble the DNM and check its size and homogeneity by native page. Then prepare 160 microliters of F-sub in the reaction buffer.
Prepare seven aliquots of 160 microliters of preassembled DNM, F-sub, and DZA in the reaction buffer. Add the analyte to tubes two to seven, and gently mix the components. Then spin down the tubes and divide the solutions into three 50-microliter portions in a black 96-well plate.
Seal the plate with an optically-transparent film. Incubate the plate in a water bath at 55 degrees Celsius for one hour. Then spin down the plate.
Afterward, measure fluorescence at 480 nanometers excitation wavelength, and 525 nanometers emission wavelength.
