African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12193

Full Length Research Paper

Leaf storage conditions and genomic DNA isolation efficiency in Ocimum gratissimum L. from Kenya

Lexa G. Matasyoh1*, Francis N. Wachira2, Miriam G. Kinyua3, Anne W. Thairu Muigai1 and Titus K. Mukiama4
1Department of Botany, Jomo Kenyatta University of Agriculture and Technology, P. O. Box 62000, Nairobi, Kenya. 2Department of Biochemistry, Egerton University, P. O. Box 536, Njoro, Nakuru, Rift Valley, Kenya. 3Kenya Agricultural Research Institute, P.O. Njoro, Nakuru, Kenya. 4Department of Botany, Nairobi University, P. O. Box 29053, Nairobi, Kenya.
Email: [email protected]

  •  Accepted: 16 January 2008
  •  Published: 04 March 2008

Abstract

Storage of plant tissues for DNA is important to avoid degradation of DNA. Preliminary studies were conducted on Ocimum gratissimum L. in order to establish the storage conditions for the collected samples before DNA extraction. Secondly, the aim was to determine the best protocol for the extraction of high quality DNA, which would later be used for molecular analysis. DNA was extracted from the samples one month after field sampling. During the DNA extraction, four protocols were used; the modified hexadecyltrimethyl ammonium bromide (CTAB) mini preparation method described by Doyle and Doyle (1990), with reductants either mercaptoethanol or dithiothreitol; the modified sodium dodecyl sulphate (SDS) mini preparation method of Edwards et al. (1991) with redundant either mercaptoethanol or dithiothreitol. The DNA was purified, treated with RNase, quantified and examined for intactness using gel electrophoresis method. Good quality and high yield DNA could only be extracted with the buffer containing the detergent SDS and the reducing agent dithiothreiotol.

 

Key words: Ocimum gratissimum L., Sodium dodecyl sulphate (SDS), hexadecyltrimethl ammonium bromide (CTAB), dithiothreitol, mercaptoethanol.