African Journal of DNA and Cell Biology Vol.  2 (4) pp. 121-125, April, 2015.  © International Scholars Journals

Full Length Research Paper

A productive DNA separation technique for tropical plants

Wen Ching-Ling,

Institute of Tropical Biosciences and Biotechnology, China Agricultural University, Beijing, China.

Email: wen.ching21@yahoo.com

Accepted 3 March, 2015

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).