【摘要】 富含花青素的马铃薯块茎因含有大量多糖和多酚类次级代谢产物而难以大量提取高质量的总RNA。本研究以紫色和红色马铃薯为材料,利用改良Trizol法实现了块茎总RNA的快速高质量提取。试验结果表明,在Trizol法中添加2%(V/V)"-巯基乙醇、0. 75%(W/V)十六烷基三甲基溴化铵(CTAB)和7. 5%(W/V)醋酸钾(KOAc),可以有效去除多糖、多酚类物质,获得高产、完整的总RNA。改良Trizol法提取时间短、操作方便,提取的RNA通过了actin RT-PCR和18S rRNA RT-q PCR检验,可以用于后续的分子生物学试验。更多还原

【Abstract】 Potato tubers rich in anthocyanins contained large quantities of polysaccharides and polyphenols,which dramatically dragged the yield and quality of total RNA extraction. Hence,we optimized the Trizol protocol for quick and high-purity total RNA extraction from purple-and red-fleshed potato tubers.The results showed that adding 2%(V/V)-mercaptoethanol,0. 75%(W/V) cetyltrimethylammonium bromide(CTAB) and 7. 5%(W/V) potassium acetate(KOAc) to Trizol method could effectively remove polysaccharides and polyphenols and obtain high-yield and intact RNA samples. The modified Trizol method had advantages of short extraction time and easy operation,and the extracted RNA had passed the actin RT-PCR and 18 S rRNA RT-qPCR tests,which could be used for subsequent molecular biological experiments.更多还原