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[Method on PCR detection of bovine materials in animal feedstuffs]
Jijuan Cao1, Xing-an Lu, Gang Wang
1Liaoning Entry-Exit Inspection and Quarantine Bureau, Dalian 116001, China.
Abstract:
The reliable, simple and easy to operate methods were established to extract DNA from animal feedstuffs. A set of PCR primers were designed for bovine specific mitochondrial DNA sequence. With this method, PCR can be successfully utilized to evaluate the presence of bovine materials in animal feedstuffs. A set of commonly used primers of 18Sr-RNA were designed as endogenuous reference gene, and check on the quality of template extraction from animal feedstuffs, and avoid the result of false negative. PCR amplification condition was one cycle of 96 degrees C for 2 min, then 35 cycle of (94 degrees C for 40s, 60 degrees C for 50s, 72 degrees C for 60s), and finally one cycle of 72 degrees C for 5 min. PCR amplified specific gene sequence 271 bp for bovine materials from animal feedstuffs.