title: A powerless on-the-spot detection protocol for transgenic crops within 30 min, from leaf sampling up to results.
abstract: The requirement of power-dependent instruments or excessive operation time usually restricts current nucleic acid amplification methods from being used for detection of transgenic crops in the field. In this paper, an easy and rapid detection method which requires no electricity supply has been developed. The time-consuming process of nucleic acid purification is omitted in this method. DNA solution obtained from leaves with 0.5 M sodium hydroxide (NaOH) can be used for loop-mediated isothermal amplification (LAMP) only after simple dilution. Traditional instruments like a polymerase chain reaction (PCR) amplifier and water bath used for DNA amplification are abandoned. Three kinds of dewar flasks were tested and it turned out that the common dewar flask was the best. Combined with visual detection of LAMP amplicons by phosphate (Pi)-induced coloration reaction, the whole process of detection of transgenic crops via genetically pure material (leaf material of one plant) could be accomplished within 30 min. The feasibility of this method was also verified by analysis of practical samples.
authors.affiliation: College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou, 310058, China.
authors.forename: Liu
authors.initials: L
authors.lastname: Wang
authors.affiliation: College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou, 310058, China.
authors.forename: Rui
authors.initials: R
authors.lastname: Wang
authors.affiliation: College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou, 310058, China.
authors.forename: Yonghua
authors.initials: Y
authors.lastname: Yu
authors.affiliation: College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou, 310058, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Zhang
authors.affiliation: Zhejiang Academy of Agricultural Science, Hangzhou, 310021, China.
authors.forename: Xiaofu
authors.initials: X
authors.lastname: Wang
authors.affiliation: College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou, 310058, China.
authors.forename: Yibin
authors.initials: Y
authors.lastname: Ying
authors.affiliation: College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou, 310058, China. wujian69@zju.edu.cn.
authors.forename: Jian
authors.initials: J
authors.lastname: Wu
authors.affiliation: Zhejiang Academy of Agricultural Science, Hangzhou, 310021, China.
authors.forename: Junfeng
authors.initials: J
authors.lastname: Xu
mesh_terms: D018556:Crops, Agricultural / Q000737:chemistry* / Q000235:genetics; D017931:DNA Primers; D016002:Discriminant Analysis; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D030821:Plants, Genetically Modified / Q000737:chemistry* / Q000235:genetics
chemical_list: D017931:DNA Primers
keywords: Crude DNA extract; Loop-mediated isothermal amplification; Transgenic crop detection; Unpowered device; Visual detection