Abstract: The present invention is related to a novel method for assaying high quantity of nucleic acid fragments, by using the single-stranded anchor nucleic acid probe with biotin and this single-stranded anchor nucleic acid hereto form double-stranded nucleic acid with the target nucleic acid fragment. To detect the target fragment does not need electrophoresis or nucleic acid sequencing but needs the antibody against double-stranded nucleic acid or the specific single-stranded reporter nucleic acid labeled with a fluorescent dye. The present invention is also related to a kit of assaying high quantity of nucleic acid fragments. A container (the multiwell plate) coating the avidin family molecule is a tool for connecting with the biotinylated target nucleic acid and subjected to the antibody reaction or fluorescence detection.