聚合酶链式反应(Polymerase chain reaction,PCR)作为一项基本的分子生物学技术,是DNA序列扩增的强有力工具.水稻作为人类最重要的粮食作物之一被广泛研究,然而由于水稻基因组序列的GC含量相对较高,而高GC含量DNA序列的PCR扩增一般比较困难,使得水稻高GC含量序列的功能研究受阻.通过使用不同的添加剂,发现相比于二甲基亚砜和甘油等,甜菜碱对于水稻高GC含量片段的PCR扩增的增强效果最明显.通过对多个高GC含量的水稻DNA片段进行PCR扩增,发现1-2 mol/L甜菜碱对于增强PCR扩增效果显著.此外,甜菜碱对于多种DNA聚合酶作用下的PCR扩增均有增强效果.%As a powerful tool for the amplification of DNA sequences,polymerase chain reaction (PCR) has been commonly used in molecular biology.Rice is one of the most important crops and has been widely studied as a model plant.However,the rice genome is rich in GC content,and the amplification of sequence with high GC content by standard PCR procedures is rather tough,which hinders the functional characterization of rice genome sequences with high GC content.To overcome this problem,several additives have been described to improve amplifications.Here after testing different additives,we found that betaine most significantly improved the amplification of rice GC-rich DNA sequences,compared to dimethyl sulfoxide and glycerol.Then we carried out PCR amplifications of multiple GC-rich rice DNA segments,and we found that the 1 mol/L to 2 mol/L betaine demonstrated remarkable effect in enhancing PCR amplification.In addition,betaine was effective in improving PCR amplifications at multiple DNA polymerases.
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