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为了确定出适用于玉米DNA指纹库构建的引物,对部分并不是完全符合玉米DNA指纹库构建引物筛选原则的引物进行重新设计与优化,使其基本符合核心引物设计的要求。以umc2105、umc1705和phi299852为例,下载原引物的基因组序列,利用两个常用的引物设计软件Primer Premier5.0和Oligo6.57进行引物的重新设计及评估,在Tm值、GC含量、引发效率、错误引发率、3’端G值、引物二聚体等方面对引物进行改善,结果表明,经过对原引物的重新设计和优化,新设计的引物均能清晰的扩增,扩增效果得到了明显的改善,并且均能在统一的条件下进行实验,符合了玉米DNA指纹库构建的要求。
In order to determine the suitable primers for the construction of DNA fingerprinting libraries, we designed and optimized some primers that are not completely consistent with the principle of primer selection for maize DNA fingerprinting, which basically conformed to the requirements of core primer design. Taking umc2105, umc1705 and phi299852 as an example, the genomic sequences of the original primers were downloaded, and two primer design software Primer Premier5.0 and Oligo6.57 were used to redesign and evaluate the primers. Tm value, GC content, elicitation efficiency, Mispriming rate, 3 ’G value, primer dimer and other aspects of the primer to improve, the results show that after redesign and optimization of the original primer, the newly designed primers can be a clear amplification, amplified results obtained Obvious improvement, and can be carried out under the uniform conditions of the experiment, in line with the requirements of the construction of DNA fingerprinting maize.