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利用特殊性引物,应用SYBR Green I实时PCR技术鉴定检测了商业化种植的转基因油菜RT73品系。结果表明,应用实时PCR技术,利用SYBR Green I染料能选择性结合双链DNA的特点,可检测到RT73品系特异性靶序列扩增所产生的荧光信号,且通过溶解曲线确定其熔点值为(80.2±0.5)℃,而对其他油菜品系则检测不到荧光信号。SYBR Green I实时PCR能通过溶解曲线有效地区分了特异性产物、非特异性产物以及引物二聚体,是基因鉴定检测的新方法。
The SYBR Green I real-time PCR technique was used to identify and test the commercially grown transgenic canola RT73 strain using specific primers. The results showed that using real-time PCR technology, SYBRGreen I dye can selectively bind to double-stranded DNA, RT73 strain can be detected by specific target amplification amplification of the fluorescence signal, and by melting curve to determine the melting point of (80.2 ± 0.5) ℃, while no fluorescence signal was detected in other rapeseed lines. SYBRGreen I real-time PCR can effectively differentiate specific products, nonspecific products and primer-dimers through the dissolution curve, which is a new method for gene identification and detection.