花生子叶RNA提取方法比较与分析  

Comparison and Analysis on Extraction Method for Total RNA from Peanut Cotyledon

作　　者：杨晨[1,2] 徐凤花[1] 单世华[2] 刘宇[2,3] 李春娟[2] 张廷婷[2] 闫彩霞[2] 

机构地区：[1]东北农业大学,黑龙江哈尔滨150030 [2]山东省花生研究所,山东青岛266100 [3]中国海洋大学,山东青岛266100

出　　处：《山东农业科学》2011年第1期1-5,共5页Shandong Agricultural Sciences

基　　金：国家自然科学基金(30771361);国家"863"计划项目(2006AA10A115)资助

摘　　要：以HY22组培苗为材料,比较改良胍酸酚抽提法、改良CTAB法及离心柱式试剂盒法提取RNA的效果,以筛选出适合花生子叶RNA提取的方法。结果表明:试剂盒法由于样品在通过离心柱时会出现阻塞现象,易使RNA提取结果受到较大影响,而改进后的胍酸酚抽提法与改良CTAB法则能避免该情况且更节约成本;经紫外分光光度法与电泳检测,改进后的两种方法的RNA提取效果均较好;RT-PCR结果表明,提取的总RNA可直接用于分子克隆和基因表达分析等分子生物学实验。To select the optimum extraction method for total RNA from peanut cotyledon, the extraction effect of modified gnanidine - acid - phenol extraction method, modified CTAB method and column centrifugation extraction kit method were compared with HY22 tissue culture seedlings as materials. The results showed that the RNA extraction effect of extraction kit method could be significantly affected because the samples could always be blocked in columns. The two modified methods could avoid this phenomenon and save cost, so they had better extraction effect by the detection of UV spectrophotometer and electrophoresis. The RT - PCR results showed that the total RNA extracted by the modified methods could be directly used for molecular biology experiments, such as molecular cloning and gene expression analysis.

关 键 词：花生 子叶 RNA提取 胍酸酚改进法 RT—PCR 

分 类 号：Q781[生物学—分子生物学]