SIMPLE DNA ISOLATION PROTOCOL FOR THE TUBEROUS CROP, Gloriosa superba L.

The isolation of intact and high-molecular mass genomic DNA is an initial and essential step for many molecular biology applications like restriction enzyme fingerprinting, polymerase chain reaction (PCR) etc. The presence of essential oils, polysaccharides, polyphenols and other secondary metabolites in the medicinal plants can hamper and reduce the yield and purity of extracted DNA. In this protocol, we describe simple modifications to the conventional cetyl trimethylammonium bromide (CTAB) method for reliable isolation of high molecular weight genomic DNA from Gloriosa superba - a potent medicinal plant. The purity of genomic DNA was confirmed by excellent absorbance (A260/A280) ratio of ~1.8 calculated from the spectrophotometric readings and the DNA concentration ranged from 75 to 308 µg/g of leaves. The PCR amplification from the isolated DNA was optimized to produce clear amplification in all the germplasm tested as mentioned in our earlier research. Besides being rapid, effective and technically easy, the protocol can also be used for the isolation of genomic DNA from other crop plants too. [ABSTRACT FROM AUTHOR]