【摘要】 甘薯块根富含多糖(主要为淀粉),提取RNA时,二者极易形成胶状沉淀而难以溶解,严重制约了甘薯块根RNA的提取产量和质量,从而影响后续分子生物学的研究。本研究以甘薯新品系农大辐14的块根为材料,采用异硫氰酸胍提取液结合10%(w/v)醋酸钾(KOAc)和1%(w/v)十六烷基三甲基溴化铵(CTAB)沉淀多糖,在1.5h内完成块根总RNA的提取,平均产量为226μg/gFW,A260/A280值为1.95。提取的总RNA可成功进行单链cDNA的合成和反转录PCR。与已有相关报道相比,该方法在保证总RNA纯度的前提下,产量有所提高,所用时间仅为前人报道的1/3￣1/10。更多还原

【Abstract】 Storage roots of sweetpotato (Ipomoea batatas (L.) Lam.) are the major organs for storing starch, a kind of polysaccharides, which could be easily combined with RNA to form a dissolving gelatinous sediments. Thus, it easily effects on the RNA isolation from storage roots of sweetpotato. In present paper we employed the storage roots of sweetpotato line Nongdafu 14 as material to establish an efficient RNA isolation protocol that only needs an hour and a half consumed. An acid thioguanidium extraction buffer combined with subsequent 10% (w/v) potassium acetate (KOAc) and 1% (w/v) cetyltrimethyl ammonium bromide (CTAB) was used to precipitate the rich polysaccharides. The outputs of RNA averaged 226 μg/g fresh weight and A260/A280 ratios were about 1.95. The RNA isolated by this method would be more competent for the first strand cDNA synthesis and reverse transcriptase PCR. The operating time of this method was significantly reduced to 1/3~1/10 of those previously reported, the yields might be increased a little and the quality such as purity would be guaranteed as well.更多还原