富含多糖的甜瓜果肉中提取总RNA方法的比较研究  

Comparative Test of Methods of Extracting Total RNA from Melon(Cucumis melo) Rich in Polysaccharides

作　　者：郝金凤[1] 哈斯阿古拉[1] 

机构地区：[1]内蒙古大学生命科学学院生物工程中心,内蒙古自治区牧草与特色作物生物技术重点实验室,内蒙古呼和浩特010021

出　　处：《北方园艺》2010年第5期15-18,共4页Northern Horticulture

基　　金：国家自然科学基金资助项目(30660111)

摘　　要：以甜瓜品种河套蜜瓜果肉为试验材料,比较了异硫氰酸胍法、CTAB法和SDS法3种RNA提取方法,结合KAc沉淀多糖和低浓度乙醇沉淀多糖2种去糖方法。结果表明:异硫氰酸胍法结合KAc沉淀多糖的去糖效果最佳,提取的RNA中28S亮度约为18S的2倍,且OD260/OD280值在1.8以上,RNA产率为64.48μg/g。经RT-PCR获得了乙烯受体基因etr1长达2.3 kb的cDNA特异性条带,说明异硫氰酸胍结合KAc沉淀多糖法从甜瓜果肉中提取的RNA质量好、产率较高、完整性好,适合于甜瓜进一步的分子生物学研究。Took melon fruit as experimental material, three extraction methods of total RNA including guanidinium isothiocyanate extraction, CTAB extraction and SDS extraction were compared, combining two＇methods of eliminating polysaccharide by KAc or low concentration ethanol, respectively. The results showed that guanidinium isothiocyanate with KAc extraction method was more capable of efficiently eliminating polysaccharide compared with the others. For this method, the brightness of 28S rRNA of obtained RNA was two times as much as that of 18S rRNA, OD260/OD280 value was above 1.8, and the RNA productivities were 64. 48μg/g. The obtained RNA was successfully used to amplify a specific 2.3 kb cDNA fragment of melon Cm-ETR1 gene by RT-PCR. These results showed that guanidinium isothiocyanate with KAc extraction method was an effective RNA extraction method for those materials affluent in polysaccharide content, and the integrity and purity of extracted total RNA could meet the demand of molecular biology experiments.

关 键 词：甜瓜 RNA 多糖 

分 类 号：Q946[生物学—植物学]