Cao Jijuan, Lu Xing-an, Wang Gang, Chen Mingsheng
Liaoning Entry-Exit Inspection and Quarantine Bureau, Dalian 116001, China.
Wei Sheng Yan Jiu. 2003 Jan;32(1):30-1.
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.
建立了可靠、简单且易于操作的从动物饲料中提取DNA的方法。针对牛特异性线粒体DNA序列设计了一组PCR引物。利用该方法,PCR可成功用于评估动物饲料中牛源性物质的存在。设计了一组常用的18Sr-RNA引物作为内源性参照基因,用于检测从动物饲料中提取模板的质量,避免出现假阴性结果。PCR扩增条件为:96℃ 2分钟1个循环,然后(94℃ 40秒、60℃ 50秒、72℃ 60秒)35个循环,最后72℃ 5分钟1个循环。PCR扩增出动物饲料中牛源性物质的特异性基因序列为271 bp。