荒漠拟步甲科昆虫总RNA的有效提取及电泳图谱分析

目的]获得高质量的总RNA是开展分子生物学研究的重要一步,由于不同生物的机体组成成分有较大差异,需要针对不同类型的物种建立各自有效的RNA提取方法。鞘翅目昆虫富含几丁质的外壳,常规方法难以获得高质量的RNA。方法]以鞘翅目拟步甲科昆虫小胸鳖甲(Microdera punctipennis)为例,采用TRIzol试剂提取分离总RNA,方法略有改进,并对总RNA的电泳谱带特征进行鉴定。结果]利用该方法从小胸鳖甲成虫中所获得的总RNA纯度较高完整性好,进一步实验证明提取的总RNA可满足RT-PCR等多种后续实验。琼脂糖凝胶电泳分析表明,其总RNA电泳谱带主要以18S带为主,28S则很弱。结论]该提取方法对其他荒漠拟步甲科昆虫,尤其是含几丁质外壳的甲虫总RNA提取具有指导作用,小胸鳖甲总RAN的电泳谱带与哺乳动物肿瘤细胞的不同。

An Effective Method for the Extraction of Total RNA from Desert Tenebrionidae Insect and the Electrophoretogram of the Total RNA

【Objective】To obtain of high - quality total RNA is the first important step in molecular biology research.However,different organisms have different body composition,which can gready influence the quality of the extracted RNA,so it is necessary to build up RNA extracting method that adapts to different organisms.Coleopteran insects have their shells rich in chitin;therefore,it is difficult to obtain high quality RNA by conventional methods.【Method】The total RAN of Microdera punctipennis(Coleoptera: Tenebrionidae) was extracted and purified by a modified method with TRIzol reagent.The integrity and purity were assessed by gel electrophoresis and spectrophotometers.The bands of the total RNA on gel electrophoresis were identified and compared with those from mammalian cells.【Result】The results showed that the extracted total RNA was pure,of high quality and no contamination,which could be used for the subsequent experiments,such as RT - PCR.The agarose gel electrophoresis showed that the 28S band was faint,while the 18S band was prominent,it is the main band of the total RAN from Microdera punctipennis.【Conclusion】This method could be effective in extracting total RNA from other desert tenebrionid insects,especially for beedes containing chitin shells.The bands of total RNA from M.punctipwnnis is different from those from mammalian cells.