The invention provides a method and a PCR (polymerase chain reaction) reagent kit for identifying DNA (deoxyribonucleic acid) barcodes of animal medicinal materials. The PCR reagent kit comprises PCR enhancers. The PCR enhancers comprise bovine serum albumin, dithiothreitol, betaine and nonidet p40. The method and the PCR reagent kit for identifying the DNA barcodes of the animal medicinal materials have the advantages that low-abundance traditional Chinese medicine samples with difficulty in amplification, particularly, samples with difficulty in identifying DNA barcodes, such as samples of buffalo horns, antelope horns, tortoise shells, carapax trionycis, pangolins and snake slough, can be effectively amplified, accordingly, the amplification success rate which is close to 100%, of DNA barcode identification techniques can be guaranteed, and DNA barcode techniques can be widely applied.