African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12213

Full Length Research Paper

Optimised cetyltrimethylammonium bromide (CTAB) DNA extraction method of plant leaf with high polysaccharide and polyphenolic compounds for downstream reliable molecular analyses

Turaki A. A.
  • Turaki A. A.
  • Natural Resources Institute, University of Greenwich, UK.
  • Google Scholar
Ahmad B.
  • Ahmad B.
  • Federal University of Birnin Kebbi, Kebbi State, Nigeria.
  • Google Scholar
Magaji U. F.
  • Magaji U. F.
  • Federal University of Birnin Kebbi, Kebbi State, Nigeria.
  • Google Scholar
Abdulrazak U. K.
  • Abdulrazak U. K.
  • Federal University of Birnin Kebbi, Kebbi State, Nigeria.
  • Google Scholar
Yusuf B. A.
  • Yusuf B. A.
  • Federal University of Birnin Kebbi, Kebbi State, Nigeria.
  • Google Scholar
Hamza A. B.
  • Hamza A. B.
  • Natural Resources Institute, University of Greenwich, UK.
  • Google Scholar


  •  Received: 14 February 2017
  •  Accepted: 28 April 2017
  •  Published: 14 June 2017

Abstract

Yam (Dioscorea species) and banana (Musa species) leaf sample contain high levels of polysaccharide and polyphenolic compounds. Extraction of good quality DNA from such leaves is usually problematic. Therefore, there is a need to extract good quality DNA in order to perform downstream DNA analysis, especially for the detection of viruses due to their low titration in some infected leaves. The cetyltrimethylammonium bromide (CTAB) DNA extraction method was selected in order to optimise DNA extraction from plant material with high level of polysaccharides. Four steps of the Lodhi et al. (1994) method were modified to be user friendly in less resource laboratories of developing countries. Yield and purity of the extracted DNAs were quantified using a NanoDrop 2000 spectrophotometer and by migration on an agarose gel after polymerase chain reaction (PCR) amplification. The results of the DNA yields and purity were in the range of 287.40±2.23 to 424.95±1.85 ng/ul and 2.10±0.05 to 2.19±0.04, respectively. Modification was found to yield DNA of reasonable quantity, quality and purity. Furthermore, the method can be used in a laboratory with less sophisticated waste disposal system.

Key words: Deoxyribonucleic acid (DNA), cetyltrimethyl ammonium bromide (CTAB), yam, banana, tomato.