摘要
由于苏铁(Cycas revoluta)叶片中含有大量的多糖多酚等次生代谢物,常规RNA提取方法很难获得优质的RNA。在常规的CTAB法中加入了硼砂和β-巯基乙醇来消除多酚和多糖的干扰,得到了一个从苏铁叶片中有效提取RNA的方法,每克鲜叶片可获得约930μgRNA。A260/280和A260/230的纳米波长的吸收比值都约为2,表明RNA的质量较好。获得的RNA可用于Northernblot和反转录PCR等分析,也说明RNA的质量比较好。此外,改进的提取方法也适合于含有次生代谢产物的其它植物,同样可以获得优质RNA。
Conventional RNA extraction methods have been shown to produce poor-quabty RNA when applied to Cycas revoluta because of abundant secondary metabolites included. With modification of the standard eetyhrimethylammonium bromide （CTAB） method by adding borax （disodium tetraborate decahydrate） and β-mereaptoethanol to eliminate the interference of polyphenol and polysaccharides, an effective protocol was developed. This modified protocol could extract high quantities of RNA from cycad leaves at about 930μg per gram of fresh weight. Both A260/230 and A260/280 ratios were around 2, indicating that RNA is of high quality. The RNA quality was confirmed by Northern blotting analysis and reverse transcription polymerase chain reaction （RT-PCR）. The modified protocol could be successfully extended to other plants containing secondary metabolites.
出处
《云南植物研究》
CSCD
北大核心
2008年第5期593-596,共4页
Acta Botanica Yunnanica
基金
The Programs of the Natural Science Foundation of Yunnan Province(Grant No.2005C0034Q)
Demonstration of Collecting & Conserving Standardization and Sharing of Important Wild Plant Germplasm Resources(Grant No.2004DKA30430,Grant No.2005DKA21006)
the Knowledge Innovation Program of Chinese Academy of Sciences
关键词
苏铁
硼砂
RNA
提取
Cycas revoluta
Borax
RNA extraction
