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.
