兼并PCR技术反应条件的探索

Studies on the PCR conditions of Degenerate Primer

[目的]探索兼并PCR技术反应的条件。[方法]依据过氧化物酶N-末端氨基酸序列设计的兼并引物,提取幼龄梨果实的DNA和RNA并将RNA反转录为cDNA,分别以DNA、cDNA和PCR产物为模板,利用PCR和琼脂糖电泳分析的方法,在不同的退火温度和引物添加量的配比下,对PCR条件进行优化。[结果]优化了PCR反应的条件,并且得到目的基因。[结论]优化后的兼并PCR技术可以更好地应用于新基因的克隆、病毒的检测等研究领域。

〔Objective〕Studies on the PCR conditions of degenerate Primer.〔Methods〕The PCR conditions were optimized by PCR and agarose gel electrophoresis at different annealing temperatures and primer addition,by designing the degenerate primers on the according of N terminal amino acid sequence of POD,and extracting the DNA and RNA from Pear fruitlets,and RNA was transcribed into cDNA.〔Results〕The conditions of PCR reaction were optimized,and the target gene was obtained.〔Conclusion〕The optimized PCR of degenerate Primer technology can be applied to the cloning of new genes,detection of viruses and other research fields.