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A powerless on-the-spot detection protocol for transgenic crops within 30 min, from leaf sampling up to results

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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.

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Acknowledgments

This work is supported by the Synergistic Innovation Center of Modern Agricultural Equipment and Technology (NZXT01201402) and National Natural Science Foundation of China (31271617). We thank the reviewers’ elaborate advice on improving the manuscript.

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Authors and Affiliations

  1. College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou, 310058, China

    Liu Wang, Rui Wang, Yonghua Yu, Fang Zhang, Yibin Ying & Jian Wu

  2. Zhejiang Academy of Agricultural Science, Hangzhou, 310021, China

    Xiaofu Wang & Junfeng Xu

Authors
  1. Liu Wang
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  2. Rui Wang
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  3. Yonghua Yu
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  4. Fang Zhang
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  5. Xiaofu Wang
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  6. Yibin Ying
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  7. Jian Wu
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  8. Junfeng Xu
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Corresponding author

Correspondence to Jian Wu.

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Wang, L., Wang, R., Yu, Y. et al. A powerless on-the-spot detection protocol for transgenic crops within 30 min, from leaf sampling up to results. Anal Bioanal Chem 408, 657–662 (2016). https://doi.org/10.1007/s00216-015-9128-x

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  • DOI: https://doi.org/10.1007/s00216-015-9128-x

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