Comparative Analysis of DNA Extraction Methods in Two Popular Varieties of Finger Millet (Eleusine coracana) from Ethiopia
Aklilu Banjaw Leza
Department of Biology, College of Natural and Computational Sciences, Dilla University, Dilla 419, Ethiopia
Sunil Tulshiram Hajare
Department of Biology, College of Natural and Computational Sciences, Dilla University, Dilla 419, Ethiopia
Nitin M. Chauhan
Department of Biology, College of Natural and Computational Sciences, Dilla University, Dilla 419, Ethiopia
*Author to whom correspondence should be addressed.
Abstract
Aims: The aim of this investigation was to standardize the genomic DNA isolation protocol to obtain high quality and quantity DNA for genomic analysis. To accomplish this task, dry leaves of finger millet which are rich in polysaccharides and polyphenols and seeds were utilized as study group.
Methodology: Two popular finger millet varieties of Ethiopia (Tadesse and Degu) were selected and obtained from Hawassa Agricultural Research Institute, Hawassa, Ethiopia. DNA isolation was carried out by two most popular and reliable methods i.e. Cetyl Trimethyl Ammonium Bromide (CTAB) and Dodecyl Trimethyl Ammonium Bromide (DTAB) method with little modifications. The quality of DNA obtained through the two methods was comparatively evaluated.
Results: The CTAB method proved its superiority over DTAB. The purity of extracted DNA was excellent as evident by A260/A280 ratio ranging from 1.76 to 1.78, which also suggested that the preparations were sufficiently free of proteins and polyphenolics/polysaccharide compounds. However, the DTAB method failed to extract sufficient quantity and quality of genomic DNA for further genomic analysis.
Conclusions: Based on our study, our protocol can be useful for other difficult cereal crops in the future.
Keywords: CTAB, DTAB, Eluesine coracana, genomic DNA