Application of Doyle and Doyle method for DNA isolation from pamelo yellow orange (Citrus maxima Merr), lime orange (C. limon) and sunkist orange (C. sinensis)

Abstract

Various species of the genus Citrus have grown naturally and were cultivated in Indonesia as a mega-biodiversity country. In recent years, the nutritional content and antioxidant activity of citrus fruits has been used to prevent of the various chronic and degenerative human diseases including potential as an antidote to viruses. Development of the potential and superiority of Indonesian oranges including Pomelo yellow orange (Citrus maxima Merr), lime orange (C. limon) and Sunkist orange (C. sinensis) are needed to be done with molecular characterization that begins with DNA isolation. The study was conducted with the aim of applying the Doyle and Doyle method for DNA isolation to all of the three types of oranges. The method for isolate DNA from plant usually use Cethyl Trimethyl Ammonium Bromide (CTAB) and Sodium Dodecyl Sulphate (SDS). The aim of this study is to isolate DNA from the orange leaves of Pomelo Yellow Orange, Lime Orange and Sunkist Orange with the Doyle & Doyle method. The results of the isolation of orange DNA were measured using NanoDrop. The results showed that the DNA of the three oranges had purity of pomelo yellow oranges 1,83, limes 1.8 and sunkist oranges 1,51. The concentrations obtained from each of the oranges were Yellow Bali Citrus 532 ng / µl, Lime Citrus 664.5 ng / µl and Sunkist Orange 888.6 ng / µl.