Isolation and Estimation of DNA Level in Coconut Leaf (Coccos Nucifera)

Abstract

The DNA Level of Coconut leaf was determined using agarose gel electrophoresis and UV-double beam spectrophotometer. 30?g leaf sample was weighed; chemical homogenization using mortar and pestle was done using lyses buffer and “Morning fresh” detergent. Whole sample was centrifuged at 10,000 rpm for 20 minutes; the supernatant was decamped into clean microcentruifuge tubes. Ethanol (500ml) was added and mixed thoroughly and incubated at room temperature for 30 minutes. DNA is insoluble in ethanol and so will appear as white precipitate at the bottom of the tube. Sample was centrifuged at 10,000 rpm for 5 minutes, thereafter the content was exposed to the atmosphere for 10 minutes to rid off remaining solvent (Ethanol). The pellets were dissolved in 50ml of TE (Tris-EDTA) buffer. The DNA (25ml) was taken and diluted in 1.75ml of TE Buffer and absorbance read at 260nm and 280nm with purity of DNA calculated followed by Electrophoresis. 25ml of the DNA sample was taken and ran on 0.8% agarose gel electrophoresis using a standard marker for 60 minutes. The analysis was done in triplicate with the ratio of absorbance at 260 and 280nm (1.79, 1.76 and 1.84) showing the purity of the DNA sample.

Keywords: Coccos nucifera, Percentage Purity, DNA Sample, TE Buffer, Agarose Gel Electrophoresis.