Molecular techniques require isolation of genomic DNA of suitable purity. High-quality genomic DNA is of the important requirements for sharp band amplicons in a Polymerase chain reaction for molecular detection of viruses on okra. The presence of mucilaginous acidic polysaccharides, polyphenols, and different secondary metabolites in okra leaves, flower, seeds, interferes with this process to a large extent as it makes DNA unmanageable in pipetting and inhibit Taq DNA polymerase activity. In this study, we reviewed improved and reliable extraction protocols which are efficient for highly purity DNA isolation from dried leaves and seeds. Several procedures have been adopted in recent years such as the modifications of conventional CTAB Doyle & Doyle (1987); increase the volume of DNA extracting buffer (1.5ml/sample), decrease sample volume (50-60mg), higher salt concentration (5M NaCl), use of polyvinylpolypyrrolidone, omission of chloroform. Other procedure employed a high concentration of Sodium dodecyl sulphate (SDS and also Potassium chloride (KCl). The yield and quality of the isolated DNA was free of contaminants, suitable for further genomic analysis through PCR, RAPD, and high-throughput DNA isolation. These procedures do not necessarily require liquid nitrogen, easy and cost-effective laboratory materials and can be carried out within a short period of time.

Molecular technique, DNA extraction, Okra, mucilage, CTAB, SDS.

biology

Presentation: oral presentation