猪肉类罐头食品中总DNA提取方法的比较  

Comparison of DNA extraction methods for canned pork

作　　者：杨彤晖[1,2] 孟镇[1,2] 钟其顶[1,2] 仇凯[1,2] 安丽艳 李志军[1,2] 

机构地区：[1]中国食品发酵工业研究院,北京100015 [2]全国食品发酵标准化中心,北京100015

出　　处：《食品与发酵工业》2015年第4期62-67,共6页Food and Fermentation Industries

基　　金：GB/T 13213-2006猪肉糜类罐头国家标准修订计划(No.20101434-T-469)支持

摘　　要：以7种猪肉类罐头食品为研究对象,比较了EE101-02 DNA提取试剂盒、SDS法、CTAB法和异硫氰酸胍法对猪肉罐头食品DNA的提取效果。以4种方法提取的总DNA为模板,根据猪线粒体基因组(mt DNA)16s r RNA基因和Cyt b基因,利用3对通用引物分别扩增片段大小为244 bp(16s r RNA)、376 bp(Cyt b)、472 bp(Cyt b)的目的片段,以评价不同方法提取的总DNA的效果。结果表明:不同提取方法提取效果差异明显——异硫氰酸胍法提取效果最好,其提取的7种罐头的总DNA均可利用16s r RNA和Cyt b通用引物分别有效扩增出244bp、376 bp目的 DNA片段,CTAB法和SDS法次之,EE101-02 DNA提取试剂盒提取效果最差。另外绝大多数猪肉类罐头样品中未能扩增获得472 bp大小的目的片段,表明猪肉类罐头产品中DNA降解严重。Four DNA extraction methods for canned pork were compared. TRANS genomic DNA kit,the SDS method,the CTAB method and the guanidine thiocyanate extraction method were applied. DNA was extracted from seven different canned pork samples. The quantity and quality of DNA extracted was evaluated by using a set of designed primers amplifying fragments of cytochrome b or 16 s r RNA of increasing size of 244bp（ 16 s r RNA）,376bp（ Cyt b）,472bp（ Cyt b）. Results show that four DNA extraction methods have significant difference in DNA extraction. Best results were obtained with the guanidine thiocyanate extraction method,which could amplify the 244 bp and376bp DNA fragments in all seven canned pork samples. The followed methods were the CTAB method and SDS method,which had better results than TRANS genomic DNA kit. Most of the DNA extracted by all four methods were failed to amplify 472 bp fragment,indicating serious DNA degradation in canned pork. This study established an effective DNA extraction method for canned pork,which laid the foundation for poultry species identification in canned food.

关 键 词：猪肉类罐头 DNA提取 方法比较 动物源鉴别 

分 类 号：TS295.1[轻工技术与工程—农产品加工及贮藏工程]