DNA的不同提取方法对转基因产品定值的影响

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。

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

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.