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为解决玉米SSR标记DNA指纹数据库构建时,N+1峰带来的指纹数据标准化采集问题,本研究对40个玉米品种鉴定核心引物进行评估,其中13对引物的扩增产物产生N+1峰。对于这些引物,通过增加PCR程序最后一步的延伸时间和下游引物5’端加一个G(鸟嘌呤)两种方法进行N+1峰的消除。结果表明:与标准中提供的程序和引物序列相比,将PCR程序最后一步的延伸时间增加至90 min,消除了9对引物扩增产物的N+1峰,下游引物5’端加一个G能消除12对引物扩增产物的N+1峰。最终提出两种消除N+1峰的方案,均能有效改善玉米标准DNA指纹数据库的图谱质量。
In order to solve the problem of standardized collection of fingerprinting data from N + 1 peak in the construction of SSR marker DNA fingerprinting database, 40 primer pairs were evaluated in this study. Among them, 13 primer pairs amplified their N + 1 peaks . For these primers, elimination of the N + 1 peak was achieved by increasing the extension time of the last step of the PCR program and adding a G (guanine) method to the 5 ’end of the downstream primer. The results showed that the extension of the last step of the PCR program was increased to 90 min compared to the programs and primer sequences provided in the standard, eliminating N + 1 peaks of 9 primer amplification products and adding a G at the 5 ’end of the downstream primer Can eliminate 12 pairs of primer amplification products N +1 peak. Finally proposed two programs to eliminate the N +1 peak, can effectively improve the quality of maize standard DNA fingerprinting database.