An efficient DNA isolation method for tropical plants

  • Qi-Xing Huang
  • Xu-Chu Wang
  • Hua Kong
  • Yun-Ling Guo
  • An-Ping Guo

Abstract

Due to interfering components such as polysacharrides, polyphenols, etc, DNA isolation from tropical plants had been challenging. We developed a safe, universal and efficient DNA extraction method, which yielded high-quality DNA from 10 tropical plants including cassava, rubber tree, banana, etc. In the extraction buffer, 2 M NaCl was used to provide a high ionic strength reaction environment, ethylenediaminetetraacetic acid (EDTA), lauroyl sarcosine (LSS) and cetyl trimethyl ammonium bromide (CTAB) could inhibit DNase activity effectively, polyvinylpolypyrrolidone (PVPP) produced a deoxidized reaction environment, and borax enhanced the precipitation of interfering compounds. Ordinary reagents like β-mercaptoethanol, chloroform and phenol were unnecessary in this protocol, which made it safe and friendly to use. PCR and EcoR I enzyme restriction digestion results show that the obtained DNA is good enough for downstream analysis. In conclusion, this protocol is expected to be a preferable DNA extraction protocol for tropical plants.

Keywords: DNA extraction, tropical plants, cetyl trimethyl ammonium bromide (CTAB)

African Journal of Biotechnology Vol. 12(19), pp. 2727-2732

Author Biographies

Qi-Xing Huang
Institute of Tropical Biosciences and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou Hainan 571101, P. R. China; Key Laboratory of Biology and Genetic Resources of Tropical Crops, Ministry of Agriculture, P. R. China
Xu-Chu Wang
Institute of Tropical Biosciences and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou Hainan 571101, P. R. China; Key Laboratory of Biology and Genetic Resources of Tropical Crops, Ministry of Agriculture, P. R. China
Hua Kong
Institute of Tropical Biosciences and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou Hainan 571101, P. R. China; Key Laboratory of Biology and Genetic Resources of Tropical Crops, Ministry of Agriculture, P. R. China
Yun-Ling Guo
Institute of Tropical Biosciences and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou Hainan 571101, P. R. China; Key Laboratory of Biology and Genetic Resources of Tropical Crops, Ministry of Agriculture, P. R. China
An-Ping Guo
Institute of Tropical Biosciences and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou Hainan 571101, P. R. China; Key Laboratory of Biology and Genetic Resources of Tropical Crops, Ministry of Agriculture, P. R. China
Published
2016-02-07
Section
Articles

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eISSN: 1684-5315