Efficient and Cost-Effective DNA Extraction Methods from Pearl Millet Leaves without Use of Liquid Nitrogen and Phenol

Abstract

DNA extraction is a very tedious process for different plant species because of the presence of secondary metabolites and huge amounts of phenol that create the difficulty of DNA isolation. In molecular biology, DNA isolation is a very crucial step and has applications viz. gene amplification, DNA restriction digestion, and gene cloning. A simple, cost-effective, and efficient method is required for the isolation of genomic DNA of Bajra. The present work uses relatively rapid, simple, cost-effective, time-saving methods for DNA isolation from both old and young leaves of Bajra genotypes without using liquid nitrogen in the extraction buffer. Using this method, a high quality and quantity of genomic DNA were obtained from 1.0 g of both old and young leaves. The total genomic DNA was extracted and amplified by employing SSR markers, which produced reproducible results. This protocol resolves the basic problems of DNA degradation, RNA contamination, and low DNA yield due to binding and/or co-precipitation with starches and complex polysaccharides.