In order to detect the DNA from highly processed medicinal donkey-hide glue Colla Corii Asini from adulterants blended or totally substituted with horse, cattle, and pig tissues, various species-specific PCR assays based on highly repetitive SINE elements were established. For horse and donkey tissues, a two-step PCR assay was adopted, including an initial detection of equine DNA based on the equine SINE ERE-1 and a further detection of horse DNA based on the horse specific satellite. The assay can detect the equine DNA and the horse DNA from binary solid glue mixtures spiked with 0.1% Colla Corii Asini and 1% horse-hide glue respectively. In addition, for bovine and porcine detection, the primer pairs were designed upon 1.711B bovine repeat and PRE-1 respectively. The developed methods can detect bovine and porcine DNA from binary solid glue mixtures spiked with 0.1% cattle-hide glue and 0.1% pig-hide glue respectively. The result shows that our developed PCR assays were useful, convenient and sensitive for Colla Corii Asini identification.