Abstract
Pearl millet is widely grown as a multi-purpose cereal grain crop for feed, fodder, fuel and mulch predominantly in the semi-arid tropics. It is highly adapted to drought, representing an essential component of the food security and livelihood of many million poor farmers. DNA extraction is unusually difficult in some plants due to the presence of secondary metabolites that interfere with DNA isolation making it very laborious and time consuming. An effective genomic DNA extraction should be simple, cost effective, with good yield and high purity. The protocol used in this study involved the extraction of genomic DNA from fresh leaves using the Sodium dodecyl sulphate (SDS) method with slight modifications including absence of use of liquid nitrogen which is difficult and expensive to obtain in the developing World. Genomic DNA obtained from the ten pearl millet samples using this procedure was good.
Highlights
Br.) which belongs to family poaceae, is a tall, warm season, annual grass widely grown for feed, fodder, fuel and mulch in more than 26 million hectares mostly in the semi-arid tropics of sub-Saharan Africa and India (FAO & ICRISAT, 1996)
Pearl millet plays a critical role in food security, because it is has the highest levels of tolerance to heat and drought among tropical cereals (Khairwal et al, 2007), Genomic DNA extraction is usually a prerequisite for molecular and forensic analysis
Different methods have been used in isolating genomic DNA, these methods include, the Cetyl Trimethyl Ammonium Bromide (CTAB) method and its modifications (Huang et al, 2000; Doyle et al, 1987), which is the most routinely used method, the Sarkosyl Nitrogen method, the Sodium dodecyl-sulfate (SDS) method, the Phenol/chloroform method and the use of kits e.g. Promega wizardTM genomic DNA purification (Hasan et al, 2009)
Summary
Different methods have been used in isolating genomic DNA, these methods include, the Cetyl Trimethyl Ammonium Bromide (CTAB) method and its modifications (Huang et al, 2000; Doyle et al, 1987), which is the most routinely used method, the Sarkosyl Nitrogen method, the Sodium dodecyl-sulfate (SDS) method, the Phenol/chloroform method and the use of kits e.g. Promega wizardTM genomic DNA purification (Hasan et al, 2009). Though these methods have been widely used and successful in isolating DNA from different plant species, they are time consuming, laborious and not cost effective especially in resource limited laboratories. The aim of this study, was to determine a genomic DNA extraction protocol suited for isolation of reasonably pure DNA in sufficient amount from pearl millet fresh leaves which is simple and rapid, without the use of expensive chemicals and specific equipment
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