Optimization of DNA isolation and PCR - RAPD methods for molecular analysis of Urginea indica Kunth.

Abstract

is the first report on development of protocol for high purity DNA isolation from the bulb tissues of Urginea indica and optimization of conditions for RAPD-PCR analysis. Cell lyses was carried out in an extraction buffer supplemented with cetyltrimethylammonium bromide (CTAB) and sodium chloride. Two emulsification-washes with phenol: chloroform: isoamyl alcohol followed by re-precipitation with salt efficiently removed high protein and polysaccharide contaminations. Purity of the isolated DNA was confirmed by restriction digestion with Bam H I, Alu I, Hind III and Eco R I. The yield of the pure DNA ranged from 120- 150ug per gram of bulb tissue. The RAPD profiling from the isolated DNA was optimized to produce scorable, clear amplicons in all populations studied.