An Effective and Low-Cost Method for Genomic DNA Extraction from Cyclanthera Pedata Species (A Nutraceutical Plant) without Liquid Nitrogen for Inter Simple Sequence Repeat Analyses

ABSTRACT: For the DNA-based study of plant species, one important steps is to obtain high-quality DNA. However, this problematic when species contains a lot polyphenols and polysaccharides. The polysaccharides interfere with activity Taq polymerase enzyme during PCR reaction thereby affecting quality Therefore, method for DNA extraction from Cyclanthera pedata has been developed. current reveals CTAB-based approach that quick, dependable, economical specifically designed obtaining genus. These are abundant in secondary metabolites polysaccharides, which makes it difficult extract effectively high yield. present protocol also excludes use expensive liquid nitrogen, cost-friendly as well. High salt concentration (1.5 M) 2% polyvinylpyrrolidone were used buffer prevent solubility extract. In addition these substances, protein-like various enzymes precipitated by ammonium acetate removed centrifugation isolation process. isolated was assessed using agarose gel electrophoresis (0.8%) quantified an A260/A280 ratio ranging 1.7 1.9, absorbance >2,which indicates free proteins, polyphenols. extracted genomic amplified ISSR primer (UBC-825) clear banding pattern observed. This standardized provides pure without nitrogen or toxic phenolic compounds. It suitable routine molecular biology assays such RAPD, SSR, restriction digestion, southern blot, cloning techniques.