DNA的不同提取方法对转基因产品定值的影响  

Influence of different extraction method of DNA on values of transgenic products

作　　者：王鹏飞[1,2] 蒋滢[1] 徐俊锋[2] 汪小福[2] 陈笑芸[2] 缪青梅[2] 李玥莹[1] 

机构地区：[1]沈阳师范大学化学与生命科学学院,沈阳110034 [2]浙江省农业科学院农产品质量标准研究所,杭州310021

出　　处：《沈阳师范大学学报（自然科学版）》2014年第2期273-276,共4页Journal of Shenyang Normal University:Natural Science Edition

基　　金：辽宁省教育厅高等学校科学研究项目(L2012387)

摘　　要：国内外非常重视转基因产品的检测工作,主要依赖于DNA的PCR方法进行检测,其检测结果可靠、稳定、灵敏度较高。尤其在转基因产品的定值实验中,荧光定量PCR的应用起到重要的作用,但是DNA的质量是保证荧光定量PCR定值准确的关键。实验以100%的克螟稻为材料,用CTAB法、商品化的Promega、Qiagen以及TIANGEN提取试剂盒,分别提取克螟稻基因组DNA,经过核酸蛋白分析仪初步测定浓度、纯度以及离子浓度后,分别以水稻内源基因PLD和克螟稻特异性转化体引物及探针实施荧光定量PCR实验,以100%克螟稻为盲样实施定值,来确定一种更适合荧光定量PCR定值实验的DNA提取方法,最终确定Qiagen和TIANGEN提取试剂盒满足荧光定量PCR对100%的克螟稻标准物质定值实验的要求。Great attention is paid to the detection of genetically modified products at domestic and abroad,these detection methods mainly depend on PCR methods,owing to its reliability,stablility and high sensitivity.Thus real-time PCR plays an important role in the experiments of GM products value.But the quality of DNA is critical factor to guarantee the accurate result in reai-time PCR assys.Therefore Genomic DNA of kemingdao is extracted by a traditional method,CTAB,with and three Extraction Kits as Promega、Qiagen and TIANGEN.Then the concentration,purity and Ion concentration are determined by ultrospec 1 100.primers and probes of endogenous gene PLD and event-specific,100％ Kemingdao（for the bland）were used in real time PCR in order to determine a extraction method of DNA,which is more fitable for real time PCR in the values of Transgenic products.Through the experiment,Qiagen and TIANGEN are more suitable for the quantitive real-time PCR.

关 键 词：克螟稻 荧光定量PCR 提取方法 

分 类 号：Q943.2[生物学—植物学]