Efficient and Cost-Effective DNA Extraction Methods from Pearl Millet Leaves without Use of Liquid Nitrogen and Phenol
Krishna Kumar Yadav
ICAR-Indian Grassland and Fodder Research Institute, Jhansi 284003, Uttar Pradesh, India.
Neha Yadav
Chandrashekhar Azad University of Agriculture and Technology Nawabganj, Kanpur 208002, Uttar Pradesh, India.
Gaurav Sharma
*
Sardar Vallabhbhai Patel University of Agricultural and Technology, Meerut, Uttar Pradesh, India and Sustainable Agriculture and Learning Foundation, Agra, Uttar Pradesh, India.
Shalini Dixit
Chandrashekhar Azad University of Agriculture and Technology Nawabganj, Kanpur 208002, Uttar Pradesh, India.
Mahendra Singh
ICAR-Indian Grassland and Fodder Research Institute, Jhansi 284003, Uttar Pradesh, India.
Ayush Yadav
ICAR-Indian Grassland and Fodder Research Institute, Jhansi 284003, Uttar Pradesh, India.
Jyoti Kushwaha
Department of Biotechnology, Bundelkhand University, Jhansi, Uttar Pradesh, India.
*Author to whom correspondence should be addressed.
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.
Keywords: DNA, PCR, grasses, DNA extraction, marker-assisted selection (MAS)