Comparative analysis and innovation of a simple and rapid method for high-quality RNA and DNA extraction of kiwifruit

被引:5
作者
Afshar-Mohammadian, Mansour [1 ]
Rezadoost, Mohammad Hossein [2 ]
Fallah, Seyyed Fatemeh [1 ]
机构
[1] Univ Guilan, Fac Sci, Dept Biol, Rasht, Iran
[2] Univ Guilan, Fac Agr Sci, Dept Biotechnol, Rasht, Iran
来源
METHODSX | 2018年 / 5卷
基金
美国国家科学基金会;
关键词
RNA and DNA extraction; RT-qPCR; Kiwifruit; Polyphenols; Polysaccharides;
D O I
10.1016/j.mex.2018.03.008
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
RNA and DNA extraction is a requirement for the study of gene expression and has an increasingly important role in genetic studies of all fleshy fruits. RNA and DNA extraction is difficult in kiwifruit due to the significant amount of polysaccharides and polyphenols compounds. So far, no commercial kit has been developed specifically for high-quality RNA and DNA extraction in kiwifruit and the common protocols for RNA extraction have poor yields. This study developed a new protocol for high quality RNA extraction in Actinidia deliciosa. According to the results, the average yield of RNA extraction of fruit and leaf of A. deliciosa was similar to 2180.7 ng/mu l (similar to 545.175 mu g/g FW) and similar to 3424.9 ng/mu l (similar to 856.225 mu g/g FW), respectively with A(260)/A(280) between 1.95 to 2.07 and A(260)/A(230) higher than 2 indicating high RNA purity. While the averages yield of RNA extraction using previous methods from kiwifruit and leaf was 23 mu g/g FW and 527 mu g/g FW, respectively. Also, the average yields of genomic DNA from kiwifruit ranged from 52 to 98 ng/mu l with A(260)/A(230) between 0.60 to 1.64 and A(260)/A(280) between 1.40 to 1.48. To our knowledge, this is the first report of a highly efficient and rapid method of RNA and DNA extraction in kiwifruit which can be used for a broad spectrum of the all fleshy fruits. (C) 2018 The Authors. Published by Elsevier B.V.
引用
收藏
页码:352 / 361
页数:10
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