Abstract
Rapid DNA extraction is a prerequisite for molecular studies. Generally, plant tissue is ground in liquid nitrogen to isolate DNA; but, liquid nitrogen is dangerous and volatile. Besides, liquid nitrogen is not always available in many developing countries. To investigate if high quality DNA could be obtained for downstream PCR analysis without liquid nitrogen, the cowpea DNA was extracted by Hexadecyl trimethyl ammonium bromide cetyl trimethylammonium bromide (CTAB) method and sodium dodecyl sulphate (SDS) method, respectively, each with three different grinding methods, including ground in liquid nitrogen, in preheated mortar and in non-preheated mortar. The DNA was compared according to their yield, purity, integrity and functionality. The results showed that high quality DNA could be obtained by three grinding methods both in CTAB method and SDS method. Without liquid nitrogen, grinding plant tissue in preheated or non-preheated mortar with extraction buffer to extract DNA is feasible. Key words: Cowpea (Vigna unguiculata), grinding method, liquid nitrogen, DNA extraction.
Highlights
Cowpea, Vigna unguiculata (L.) Walp, Leguminosae (2n = 2x = 22), is an essential food crop in developing countries of the tropics and subtropics, especially in subSaharan Africa, Asia, and Central and South America (Singh et al, 1997), with an annual production of more than five million metric tons world-wide (FAO, 2010)
To investigate if high quality DNA could be obtained for downstream polymerase chain reaction (PCR) analysis without liquid nitrogen, the cowpea DNA was extracted by Hexadecyl trimethyl ammonium bromide cetyl trimethylammonium bromide (CTAB) method and sodium dodecyl sulphate (SDS) method, respectively, each with three different grinding methods, including ground in liquid nitrogen, in preheated mortar and in non-preheated mortar
The results showed that high quality DNA could be obtained by three grinding methods both in CTAB method and SDS method
Summary
As new tools that dramatically enhanced the efficiency of plant breeding, DNA based molecular markers have been widely used in cowpea genetic analysis (Zannou et al, 2008; Malviya et al, 2012; Badiane et al, 2012), genetic map construction (Muchero et al, 2009; Xu et al., 2011; Ouedraogo et al, 2002), QTL analysis (Muchero et al, 2010; Andargie et al, 2011; Kongjaimun et al, 2012) etc. Rapid DNA extraction with expectable quality is the prerequisite for molecular studies. The procedure is to grind plant tissue in liquid nitrogen and transfer it to a preheated extraction buffer (Dellaporta et al, 1983; Saghai-Maroof et al, 1984)
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