A Protocol for Extraction of High-quality RNA and DNA from Peanut Plant Tissues
A Protocol for Extraction of High-quality RNA and DNA from Peanut Plant Tissues
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DOI：
10.1007/s12033-011-9391-9
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发表时间：
2011-10-01
影响因子：
2.6
通讯作者：
Cui, Dangqun
中科院分区：
文献类型：
--
作者：
Yin, Dongmei;Liu, Haiying;Cui, Dangqun
花生是越来越重要的全球食物来源。然而，直到最近，缺乏有效的协议，用于提取核酸的分子研究花生发育和成熟的困难。在这里，我们描述了一种从花生组织中分离高质量RNA和DNA的方法，并成功地将这种方法应用于花生植物的根、茎、叶、花和种子。分光光度分析表明，每100毫克花生材料的总RNA平均产量为24.52 ~ 74.6 μ g，相同组织的基因组DNA平均产量为23.47 ~ 57.68 μ g。使用该方案，我们获得了1.9和2.0之间的OD 260/280值，并分离了可以以适合于RT-qPCR和表达分析的方式逆转录的RNA。此外，使用这种方法分离的基因组DNA产生可靠的限制性内切酶消化模式，并可用于Southern印迹杂交。
Peanuts are an increasingly important global food source. However, until recently the lack of effective protocols for the extraction of nucleic acids has made molecular studies of peanut development and maturation difficult. Here, we describe a method to isolate high-quality RNA and DNA from peanut tissue and have successfully applied this method to peanut plant roots, stems, leaves, flowers, and seeds. Spectrophotometric analysis showed that the average yields of total RNA from 100 mg of peanut materials ranged from 24.52 to 74.6 mu g, and those of genomic DNA from the same tissues ranged from 23.47 to 57.68 mu g. Using this protocol, we obtained OD260/280 values between 1.9 and 2.0 and isolated RNA which could be reverse transcribed in a manner suitable for RT-qPCR and expression analysis. In addition, genomic DNA isolated using this method produced reliable restriction enzyme digestion patterns and could be used for Southern blot hybridization.