Evaluating the efficiency of ethanol precipitation method in purification of gDNA and PCR product

Abstract

A numerous clean-up methods of nucleic acid were developed to achieve the requirements of downstream reactions like PCR and sequencing. The methods were varied in their mechanism, efficiency of purification and final product yield. The present study evaluated the efficiency of ethanol-sodium acetate (EOH-NaOAc3) precipitation method in purification of nucleic acid to satisfy downstream reactions requirements. The yield and purity of nucleic acid were considered as the main standard parameters to estimate the efficiency of method. Geneaid gel extraction kit DF100 was considered as a standard method for comparison. The results of methods comparison revealed that EOH-NaOAc3 method was significantly (P=0.000) surpassed the kit method in the yield of purified PCR product (93.24 and 18.37 ng/µl) with no significant differences (P=0.239) in quality (Absorbance (A260/280+) = 1.816 and 1.843) respectively. To determine the productivity of EOH-NaOAc3 method, a specific amount of genomic DNA (G-DNA) (187.93 ng/ µl) was processed and the results showed that EOH-NaOAc3 method was efficiently conserved 89.6% of total processed G-DNA (168.51 ng/µl) accompanied by significant (P=0.03) elevation of DNA purity (A260/280, 3.07 – 2.53).