African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12487

Full Length Research Paper

An efficient DNA isolation method for tropical plants

Qi-Xing Huang1,2, Xu-Chu Wang1,2, Hua Kong1,2, Yun-Ling Guo1,2 and An-Ping Guo1,2*
1Institute of Tropical Biosciences and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou Hainan 571101, P. R. China. 2Key Laboratory of Biology and Genetic Resources of Tropical Crops, Ministry of Agriculture, P. R. China.  
Email: [email protected]

  •  Accepted: 03 May 2013
  •  Published: 08 May 2013

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.

 

Key words: DNA extraction, tropical plants, cetyl trimethyl ammonium bromide (CTAB).