摘要
本研究分别利用TRIzol法、异硫氰酸胍法、改良的CTAB法及试剂盒这4种方法提取魔芋叶片的总RNA,并用紫外光谱和琼脂糖凝胶电泳对各个提取效果进行鉴定。反复试验的结果表明:TRIzol法难以提取魔芋叶片总RNA,异硫氰酸胍法提取的总RNA浓度低、杂质较多且有部分降解,这两种方法不适宜用于魔芋叶片总RNA提取;试剂盒方法提取的总RNA产率和纯度较高,但稳定性差、成功率低且成本较高;改良的CTAB法利用PVP和β-巯基乙醇去除酚类物质,水饱和酚-氯仿去除蛋白质,醋酸钾去除多糖,该方法提取得到的总RNA的纯度较高,OD260/OD280值为1.893,而且效果稳定,有良好的可重复性。因此,我们认为改良的CTAB法是一种简便、快速且经济的提取魔芋叶片总RNA的方法。
In this research, we employed four RNA extraction methods which contains TRIzol method, guanidiniurn isothiocyanate method, RNAprep pure kit method, and modified CTAB method to extract total RNA from leaves of konjac, and the samples of extracting total RNA of the four methods were tested by using ultraviolet spectrophotometer analysis and gel electrophoresis. The results of numerous repetitions experiments showed that it was difficult to isolate total RNA from leaf of konjac by using TRIzol method, and total RNA could be extracted by using guanidinium isothiocyanate method with low concentration, impurity and degradation in some degrees, so both of TRIzol method and guanidinium isothiocyanate method weren＇t suitable to extract total RNA from leaves of konjac. The higher yields and quality RNA could be obtained via RNAprep pure kit method, but this method was instable, cost higher than other methods and without enough achievement ratio. The modified CTAB method was by adding PVP and β-mercaptoethanol to remove polyphenol compounds, the mixture of chloroform and phenol to eliminate protein, KAC to separate polysaccharides, and not only the purity of RNA isolated by using modified CTAB method was higher than other extraction methods with OD260/OD280 value of 1.893, hut also this method has stable performance and good repeatability. For above reasons, modified CTAB method could be regarded as a rapid and convenient method for the RNA isolated from leaves of konjac which rich in polysaccharides and polyphenol compounds.
出处
《分子植物育种》
CAS
CSCD
2010年第1期196-200,共5页
Molecular Plant Breeding
关键词
魔芋
RNA抽提
TRIZOL法
异硫氰酸胍法
试剂盒法
改良CTAB法
Konjac （A morphophollus konjac）, RNA isolation, TRIzol method, Guanidinium isothiocyanate method,RNAprep pure kit method, Modified CTAB method
