改良热硼酸法高效提取苹果果实RNA

苹果果实、尤其是成熟期的果实中多糖和其他次生物质含量高,难以提取RNA。对此,在原热硼酸法的基础上进行了改进,通过添加提取辅助剂并根据辅助剂特性调整提取步骤,高效、稳定地从苹果果实、尤其是后期果实中提取到了高质量的RNA。所得RNA的A260/A230大于2.0,A260/A280为1.8-2.1,并且RNA产量高,其中前期果实RNA产率在13.40μg/g以上,后期果实RNA产率在7.02μg/g以上,完全可以满足RT-PCR、cDNA-AFLP等分子生物学实验的要求。

Effective extraction of total RNA in apple flesh with improved hot borate method

Apple fruit,especially ripe fruit,contain many polysaccharides and other secondary metabolites. Therefore, it is very difficult to extract RNA from ripe apple fruit. In order to resolve this problem,an improved hot berate procedure was developed for extracting high quality RNA from ripe apple fruit by using assistant agents and rearranging extraction steps. This modified method was so effective to produce ca. 13.40 μg/g total RNA from fruit at early stage, i.e., approximately 7 days post-anthesis, and ca. 7.02 μg/g from fruit at late stage, i.e. 7 days before ripening. The A260/A230 of yielded RNA was more than 2.0 while A260/A280 ranged from 1.8 to 2.1. The RNA can be applied to RT-PCR and cDNA-AFLP analysis.