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African Journal of Biotechnology

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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



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