ISWC OpenIR  > 水保所科研产出--SCI  > 2017--SCI
A universal method for direct PCR amplification of plant tissues
Li, Yuping1,2; Zhao, Huanhuan2; Yan, Xuefen2; Li, Meng2; Chen, Peng2; Zhang, Suiqi1,2; Zhang, SQ (reprint author), Northwest A&F Univ, State Key Lab Soil Eros & Dryland Farming Loess P, Yangling 712100, Shaanxi, Peoples R China.; Chen, P; Zhang, SQ (reprint author), Northwest A&F Univ, Coll Life Sci, Yangling 712100, Shaanxi, Peoples R China.
SubtypeArticle
2017
Source PublicationANALYTICAL METHODS
ISSN1759-9660
description.correspondentemailpengchen@nwsuaf.edu.cn ; sqzhang@ms.iswc.ac.cn
Volume9Issue:11Pages:1800-1805
AbstractPCR 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.
Subject AreaChemistry ; Food Science & Technology ; Spectroscopy
DOI10.1039/c6ay03156k
URL查看原文
Indexed BySCI
Publication PlaceCAMBRIDGE
Language英语
WOS IDWOS:000398435000016
PublisherROYAL SOC CHEMISTRY
Funding OrganizationNational 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]
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Cited Times:2[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.iswc.ac.cn/handle/361005/8018
Collection水保所科研产出--SCI_2017--SCI
Corresponding AuthorZhang, SQ (reprint author), Northwest A&F Univ, State Key Lab Soil Eros & Dryland Farming Loess P, Yangling 712100, Shaanxi, Peoples R China.; Chen, P; Zhang, SQ (reprint author), Northwest A&F Univ, Coll Life Sci, Yangling 712100, Shaanxi, Peoples R China.
Affiliation1.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
Recommended Citation
GB/T 7714
Li, Yuping,Zhao, Huanhuan,Yan, Xuefen,et al. A universal method for direct PCR amplification of plant tissues[J]. ANALYTICAL METHODS,2017,9(11):1800-1805.
APA Li, Yuping.,Zhao, Huanhuan.,Yan, Xuefen.,Li, Meng.,Chen, Peng.,...&Zhang, SQ .(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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