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Accepted Manuscript

A modified SDS-based DNA extraction method for raw soybean

Yimiao Xia, Fu-sheng Chen, Yan Du, Chen Liu, Guanhao Bu, Ying Xin, Boye Liu
Bioscience Reports Jan 15, 2019, BSR20182271; DOI: 10.1042/BSR20182271
Yimiao Xia
Henan University of Technology, Zhengzhou, China
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Fu-sheng Chen
Henan University of Technology, Zhengzhou, China
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  • For correspondence: fushengc@haut.edu.cn
Yan Du
Henan University of Technology, Zhengzhou, China
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Chen Liu
Henan University of Technology, Zhengzhou, China
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Guanhao Bu
Henan University of Technology, Zhengzhou, China
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Ying Xin
Henan University of Technology, Zhengzhou, China
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Boye Liu
Henan University of Technology, Zhengzhou, China
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Abstract

Soybean is the most important genetically modified (GM) oilseed worldwide. Regulations relating to the approval of biotech soybean varieties and product labeling demand accurate and reliable detection techniques to screen for GM soya. High quality extracted DNA is essential for DNA-based monitoring methods. Thus, four widely used protocols (SDS, CTAB, DP305, and DNeasy Plant Mini Kit) were compared in this study to explore the most efficient DNA extraction method for raw soya matrix. The SDS-based method showed the highest applicability. Then crucial factors influencing DNA yield and purity, such as SDS lysis buffer component concentrations and organic compounds used to isolate DNA, were further investigated to improve the DNA obtained from raw soybean seeds, which accounts for the innovation of this work. As a result, lysis buffer (2% SDS (w/v), 150 mM NaCl, 50 mM Tris-HCl, 50 mM EDTA, pH 8.0) and organic reagents including chloroform/isoamyl alcohol (24: 1, v/v), isopropanol, and ethanol corresponding to the extraction and first and second precipitation procedures, respectively, were used in the optimized SDS method. The optimized method was verified by extracting approximately 2020-2444 ng DNA/mg soybean with A260/280 ratios of 1.862-1.954 from five biotech and non-biotech soybean varieties. Only 0.5 mg of soya was required to obtain enough DNA for PCR amplification using the optimized SDS-based method. These results indicate that the screening protocol in this study achieves the highest suitability and efficiency for DNA isolation from raw soya seed flour.

  • Soybean
  • DNA extraction
  • SDS method
  • GMO detection
  • ©2019 The Author(s)

This is an Accepted Manuscript; not the final Version of Record. You are encouraged to use the final Version of Record that, when published, will replace this manuscript and be freely available under a Creative Commons licence.

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A modified SDS-based DNA extraction method for raw soybean
Yimiao Xia, Fu-sheng Chen, Yan Du, Chen Liu, Guanhao Bu, Ying Xin, Boye Liu
Bioscience Reports Jan 2019, BSR20182271; DOI: 10.1042/BSR20182271
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A modified SDS-based DNA extraction method for raw soybean
Yimiao Xia, Fu-sheng Chen, Yan Du, Chen Liu, Guanhao Bu, Ying Xin, Boye Liu
Bioscience Reports Jan 2019, BSR20182271; DOI: 10.1042/BSR20182271

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Keywords

Soybean
DNA extraction
SDS method
GMO detection

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