›› 2011, Vol. 38 ›› Issue (3): 84-86.

• 生物技术 • 上一篇    下一篇

T-RFLP快速鉴定生鲜肉中牛羊成分的研究

田金辉1,李宝明2,尉婷媛2,周占琴1,武会娟2   

  1. (1.西北农林科技大学动物科技学院,陕西杨凌 712100; 2. 北京市理化分析测试中心,北京 100094)
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2011-03-20 发布日期:2011-03-20
  • 通讯作者: 武会娟

Detection of Cattle-derived and Sheep-derived Component in Fresh Meat by T-RFLP Method

TIAN Jin-hui1,LI Bao-ming2,WEI Ting-yuan2,ZHOU Zhan-qin1,WU hui-juan2   

  1. (1.College of Animal S&T,Northwest A&F University,Yangling 712100,China;2.Beijiing Centre for Physical & Chemical Analysis,Beijing 100094,China)
  • Received:1900-01-01 Revised:1900-01-01 Online:2011-03-20 Published:2011-03-20
  • Contact: WU hui-juan

摘要: 试验旨在建立快速检测生鲜肉中牛、羊成分的定性分析方法。通过T-RFLP法,上游引物的5′端用6-FAM进行荧光标记,下游引物5′端用ROX进行荧光标记,PCR结束后选择限制性内切酶HinfⅠ、BlnⅠ进行酶切消化,在测序仪上通过毛细管电泳即可检测荧光标记的两个末端的限制性片段。利用不同物种产生的不同峰谱达到对物种快速定性的目的。该法能够对生鲜肉中牛、羊肉进行定性鉴定分析,不同物种产生不同峰谱,限制性内切酶HinfⅠ酶切牛肉样品所产生的限制性末端片段长度为46和373 bp,限制性内切酶BlnⅠ酶切羊肉样品所产生的限制性末端片段长度为203和515 bp。

关键词: COⅠ; PCR; 末端限制性片段长度多态性; 牛;

Abstract: To establish the qualitative analysis ways of fast testing the components of fresh sand mutton, taking strong measures against adulterations in the fresh meet. Using the T-RFLP method, the forward PCR primer was labeled with 6-carboxyfluorescein amino hexy(6-FAM) fluorescent dye and reverse PCR primer was labeled with ROX fluorescent dye after digestion by restriction enzyme HinfⅠand BlnⅠ. The restricition fragment were analysed by capillary electrophoresis using an antomated DNA sequencer. We can realize species identification through their different peak. Qualitative species identifying in the fresh beef and mutton were successfully identified by this rapid molecular technique. The PCR products of beef DNA which were digested with Hinf Ⅰ demonstrated 46 and 373 bp, meanwhile 203 and 515 bp for goat DNA which were digested with BlnⅠ.

Key words: COⅠ; PCR; terminal restriction fragment length polymorphism; cattle; goat

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