冰冻切片法破碎大鼠心肌组织提取RNA的实验研究

目的:通过比较液氮研磨法和冰冻切片法破碎大鼠心肌组织来提取RNA的效果,证明利用冰冻切片机破碎动物组织提取RNA是一种简便实用的方法。方法:用液氮研磨法和冰冻切片法破碎大鼠心肌组织并提取RNA,通过RNA浓度、纯度及完整性的检测,以及扩增β-actin基因来比较两种方法的效果。结果:液氮研磨法和冰冻切片法提取的RNA浓度分别为2.59μg/μl、2.05μg/μl。琼脂糖凝胶电泳检测可见两条明显的条带。28s rRNA与18s rRNA条带亮度之比大于1。β-actin基因的PCR扩增在约445bp处有与预期长度一致的明亮条带。结论:本实验结果显示,冰冻切片法破碎心肌组织能达到液氮研磨法的效果,是一种成本低,效果好,快速简便的破碎动物组织提取RNA的方法。

Research on frozen section method for breaking rat myocardial tissue to extract RNA

objective: To compare liquid nitrogen grinding method with frozen section method for breaking rat myocardial tissue to extract RNA and to demonstrate that the latter is a simple and effective way.Methods: Rat myocardial tissue was broken by liquid nitrogen grinding method and frozen section method,and RNA was extracted.Then the RNA concentration,purity and integrity were detected.Eventually the RNA was used as template for the amplification of f-actin gene by RT-PCR.Results: The RNA concentration extracted by Liquid nitrogen grinding method and frozen section method was 2.59μg/μl,and 2.05μg/μl respectively.Agarose gel electrophoresis exhibit two distinct bands apparently.The brightness ratio of the 28s rRNA and the 18s rRNA bands was greater than 1.β-actin gene amplified by RT-PCR had bright bands of expected length in 445bp.Conclusion: The experimental results show that frozen section method can achieve the same effect as liquid nitrogen grinding method.Besides,it is a less expensive,effective and simple method for breaking animal tissue to extract RNA.