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A universal method for direct PCR amplification of plant tissues

文献类型：期刊论文

作者Li, Yuping1,2; Zhang, SQ (reprint author), Northwest A&F Univ, Coll Life Sci, Yangling 712100, Shaanxi, Peoples R China.; Chen, P; Zhang, SQ (reprint author), Northwest A&F Univ, State Key Lab Soil Eros & Dryland Farming Loess P, Yangling 712100, Shaanxi, Peoples R China.; Zhang, Suiqi1,2; Chen, Peng2; Li, Meng2; Yan, Xuefen2; Zhao, Huanhuan2
刊名ANALYTICAL METHODS
出版日期2017
卷号9期号:11页码:1800-1805
ISSN号1759-9660
DOI10.1039/c6ay03156k
文献子类Article
英文摘要PCR is a vital tool in modern biology; however, it can be costly owing to the price of commercial DNA purification kits. DNA purification is time consuming and rare material used for DNA template purification during transgenic mutant screening can be risky. There is, therefore, an urgent need to develop alternative approaches. Here, we describe a convenient and efficient method for direct PCR amplification of plant tissues. In this method, plant tissue samples are obtained using micropipettes and, after incubation with a casein alkaline solution, are used directly as DNA templates for PCR. In addition, 20 mM ammonium sulfate and a high-fidelity DNA polymerase fused to sso7d (a small DNA-binding protein) are essential components of this system. We applied this method to tartary buckwheat, which is rich in secondary metabolites, and we found this method to be effective in maize, wheat, Arabidopsis, and tobacco. All the steps of the protocol can be carried out on a thermal cycler. Moreover, the minor injuries to the plants when collecting samples have no effect on their growth and survival. Our new protocol offers a considerable simplification of present direct PCR approaches and it will be particularly useful for screening transgenic mutants and trace amounts of precious materials.
学科主题Chemistry ; Food Science & Technology ; Spectroscopy
URL标识查看原文
出版地CAMBRIDGE
语种英语
WOS记录号WOS:000398435000016
出版者ROYAL SOC CHEMISTRY
资助机构National Natural Science Foundation of China [31171606, 30400282]; Basic Research Fund of the Northwest AF University [QN2009064] ; National Natural Science Foundation of China [31171606, 30400282]; Basic Research Fund of the Northwest AF University [QN2009064] ; National Natural Science Foundation of China [31171606, 30400282]; Basic Research Fund of the Northwest AF University [QN2009064] ; National Natural Science Foundation of China [31171606, 30400282]; Basic Research Fund of the Northwest AF University [QN2009064]
源URL[http://ir.iswc.ac.cn/handle/361005/8018]  
专题水保所科研产出--SCI_2017--SCI
通讯作者Zhang, SQ (reprint author), Northwest A&F Univ, Coll Life Sci, Yangling 712100, Shaanxi, Peoples R China.; Chen, P; Zhang, SQ (reprint author), Northwest A&F Univ, State Key Lab Soil Eros & Dryland Farming Loess P, Yangling 712100, Shaanxi, Peoples R China.
作者单位1.Northwest A&F Univ, State Key Lab Soil Eros & Dryland Farming Loess P, Yangling 712100, Shaanxi, Peoples R China
2.Northwest A&F Univ, Coll Life Sci, Yangling 712100, Shaanxi, Peoples R China
推荐引用方式
GB/T 7714		 Li, Yuping,Zhang, SQ ,Chen, P,et al. A universal method for direct PCR amplification of plant tissues[J]. ANALYTICAL METHODS,2017,9(11):1800-1805.
APA Li, Yuping.,Zhang, SQ .,Chen, P.,Zhang, SQ .,Zhang, Suiqi.,...&Zhao, Huanhuan.(2017).A universal method for direct PCR amplification of plant tissues.ANALYTICAL METHODS,9(11),1800-1805.
MLA Li, Yuping,et al."A universal method for direct PCR amplification of plant tissues".ANALYTICAL METHODS 9.11(2017):1800-1805.

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来源：水土保持研究所

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