摘要
为了获得高质量的枣RNA,本研究从所得RNA的纯度与浓度、操作时间和成本等方面比较了Trizol试剂法、传统CTAB法、热硼酸法、试剂盒法以及改良CTAB法5种方法提取枣不同组织总RNA的效果,并筛选出了最适合提取枣不同组织样品RNA的具体方法。结果表明:(1)在本研究试验的5种提取方法中,枣不同的组织RNA提取需要不同的方法。枣组培苗RNA提取的最佳方法是热硼酸法,枣芽和幼叶是试剂盒法,花和果实需要用改良CTAB法。(2)通过正交试验,得出枣树花和果实RNA提取适宜的改良CTAB法为在原有CTAB法的基础上水浴20 min、加入Na Ac的p H值为4.8、加入2倍体积的异丙醇。(3)枣RNA提取过程中的其他条件包括:RNA储存液保存的材料提取出的RNA质量最高,使用TBE缓冲液替代TAE进行电泳可以提高RNA的电泳亮度,4℃低温离心更有利于组培苗和芽的总RNA的提取,而叶片、花以及果实的RNA提取室温即可。
For high-qu ality juju be RNA, this study compared the Trizol method, traditional CTAB method, hot borate method, kits and modified CTAB method 5 ways from the RNA purity and concentration, operating time and cost to extract total RNA from different tissues of Chinese date. The results show that:(1) In the five kinds of extraction method, different organizations correspond to different extraction methods. Hot borate method was the optimal method to isolate RNA from tissue-culture plantlets; kit method was suitable to isolate RNA from buds and leaves, modified CTAB method is a common method to isolate RNA from flowers and fruits.(2) Orthogonal experiment result that modified CTAB method, on a water bath 20 min, added Na Ac p H of 4.8, 2 times volumes of isopropanol, was more suitable to extraction RNA in jujube's flower and fruit based on the original CTAB method.(3) Other conditions on Chinese date's RNA extraction include, extracted RNA from the material that save in RNA storage solution can have highest quality RNA. Electrophoresis using TBE buffer can improve the brightness of the RNA electrophoresis. 4℃ low temperature is more conducive to total RNA was extracted by centrifugation of tissue culture and shoots, flowers and fruits of RNA extraction from leaves suit to room temperature.
出处
《分子植物育种》
CAS
CSCD
北大核心
2015年第12期2871-2882,共12页
Molecular Plant Breeding
基金
"十二五"国家科技支撑项目(2013BAD14B0302)
国家自然科学基金(31400578)共同资助
