The effects of three different grinding methods in DNA extraction of cowpea (Vigna unguiculata L. Walp)

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.

Keywords: Cowpea (Vigna unguiculata), grinding method, liquid nitrogen, DNA extraction

African Journal of Biotechnology Vol. 12(16), pp. 1946-1951