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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。
The CTAB lysis buffer samples were routinely incubated and sonicated during the DNA extraction process with rice seeds of 0.1% (m / m) and 0.01% (m / m) contents of transgenic rice Kefeng No.6. Processing for comparative analysis. The results showed that the concentration of DNA in CTAB buffer solution was significantly higher than that in routine incubation at 10 min, 30 min or 1 h incubation time. The transgene The differences of Ct values between the components NOS, Cry1Ab and Kf6 (Ub-C) detected by fluorescence were very significant. Ultrasonic incubation for 30 min and 1 h could basically achieve the results of fluorescent PCR detection of samples incubated 4 h or 8 h by routine incubation, Which makes the detection time of fluorescent PCR of rice seed gene at least 3 ~ 7 h, which greatly improves the detection efficiency.