The invention provides a kit and a usage method thereof. The kit comprises one or more pairs of 16 upstream and downstream primer pairs for amplifying STR loci, namely, D3S1358, D5S818, D7S820, D8S1179, D13S317, D16S539, D18S51, D21S11, CSF1PO, FGA, TH01, TPOX, vWA, Penta E, Penta D and Amelogenin, wherein the sequences of the 16 pairs of upstream and downstream primers are shown as SEQ ID NO:1 to SEQ ID NO:32. By adopting the kit and the usage method provided by the invention, the related 16 pairs of upstream and downstream primers have the advantages of high specificity and high amplification efficiency, and genotyping of the 16 STR loci can be performed rapidly and accurately.