A Comprehensive Method to Isolate High Quality DNA from the Cultivars of Hibiscus

Abstract

An attempt was made to develop the best method to extract DNA from four selected varieties of the ornamental and medicinal plant, Hibiscus rosa-sinensis which can be distinguished one from the other morphologically. DNA extraction was carried out in several sets and each time there were more modifications that have been documented to bring out an efficient method to extract DNA from this plant of the Mavlacea family, which have a high content of gummy, mucilaginous substances. That apart, the DNA extracted was qualified and quantified based on the Beer-Lambert's law using UV-Visible Spectrophotometer. The results were not up to the mark of purity but further modifications can be made in this respect. Then a comprehensive protocol to isolate the DNA from Hibiscus sp was developed. The plant genomic DNA was extracted and also restriction digested by several combinations of restriction enzymes and conditions of incubation. The best one was found to completely digest the genomic DNA and this was the combination of three restriction enzymes Eco RI, Bam HI, Hind III at 37ºC for 15-18 hours. The dual success in developing a standardized condition for restriction digestion of genomic DNA from this species as well as the obtaining of completely digested plant genomic DNA can be further used in DNA Analysis of the related cultivars, and their RAPD analysis can be carried out which helps to identify and characterise the plants within this species as part of germplasm conservation.

Comparison of the efficiency of some of the most usual DNA extraction methods for woody plants in different tissues of <em>Vitis vinifera</em> L.

“…To be more specific, an efficient extraction and purification protocol must 1) provide pure, intact and functional DNA, 2) give a good yield, 3) be fast and simple, 4) be economic and 5) be reliable (Reddy, 2009). Several methods for the extraction and purification of DNA from woody plants that are theoretically appropriate for Vitis vinifera have been described in the literature.…”

Research Article A simple and inexpensive modified CTAB and silica particles-based method for recalcitrant plant genomic DNA isolation

“…Several methodologies to isolate genomic DNA from plants have been reported; however, there are few reports related to obtaining sufficient quantities of high-quality DNA for molecular work from recalcitrant species (Reddy, 2009;Azmat et al, 2012;Alexander 2016;Rezadoost et al, 2016;Arruda et al, 2017;Swetha et al, 2018), which contain comparatively high quantities of polysaccharides and secondary metabolites, posing problems in downstream applications of the DNA.…”