超声波处理对水稻种子转基因成分荧光PCR检测的影响

Effect of ultrasonic treatment of rice seed samples to real-time fluorescence PCR detection of genetically modified ingredients

以转基因水稻科丰6号含量0.1%（m/m）与0.01%（m/m）的水稻种子为样品,对影响检测结果的DNA提取过程中样品CTAB裂解缓冲液常规温育与超声波温育处理进行比较分析。结果表明,在不同样品颗粒细度的样品中,无论温育时间10 min、30 min还是1 h,CTAB缓冲液超声波温育处理与常规温育处理比较,获得的DNA质量浓度有明显提高,转基因成分NOS、Cry1Ab、Kf6（Ub-C）的荧光PCR检测Ct值差异达到极显著,超声波温育处理30 min、1 h可基本达到常规温育处理4 h或8 h的样品荧光PCR检测效果,使得水稻种子转基因成分荧光PCR检测时间至少节省3-7 h,大幅度提高了检测效率。

Rice seed containing transgenic rice kefeng 6 of 0.1%（m/m） and 0.01%（m/m） used as samples,the effect of ultrasonic treatment of hexadecyltrimethyl ammonium bromide（CTAB） lysis buffer to incubate rice seed samples to real-time fluorescence PCR detection of genetically modified ingredients was analyzed and compared those with regular CTAB buffer incubation. Results show that in the samples with grain different fineness,regardless of the temperature for 10 min,30 min and 1 h,DNA yields have increased significantly,the Ct values of fluorescence PCR detection for genetically modified ingredients（NOS,Cry1 Ab,Kf6（Ub-C）） show extremely significant differences between using CTAB buffer incubations with ultrasonic treatment and conventional treatment（without ultrasonic）. The effect of fluorescent PCR detection with ultrasonic incubation for 30 min and 1 h could achieve to those of conventional incubating 4 h or 8 h,to make detection for rice seeds genetically modified ingredients save at least 3 to 7h,greatly improving the detection efficiency.