Isolation of genomic DNA from groundnut plant by modified, rapid and efficient protocol

Abstract

DNA isolation is prerequisite to study the crop at molecular level. However, DNA (Deoxyribonucleic Acid) isolation is difficult in groundnut due to presence of large amount of polyphenols and polysaccharides. The age and growth stage of the plant also affect the DNA purity during isolation. In this study, newly expanded leaves and stems of five days old seedlings and matured leaves of 20 days old seedlings were used for optimization of extraction protocol. High quality DNA was obtained from all plant materials by this modified CTAB (Cetyltrimethylammonium bromide) protocol without separate purification. Optimized protocol was used to isolate DNA from newly expanded leaves of 5 days old seedlings of 100 groundnut genotypes. DNA obtained ranged from 6.3-22.1 µg/ g of sample when quantified by Nano Drop spectrophotometer and was able to amplify with Simple sequence repeats (SSR) primers during Polymerase Chain Reaction (PCR).