A New Method for Genomic DNA Extraction from Sclerotia of Sclerotium rolfsii Inciting Collar Rot of Lentil for Genomic Investigations

Abstract

Background: The prevailing methods for the genomic DNA extraction of Sclerotium rolfsii from its mycelium mat is often time consuming and yields poor quality and quantity of genomic DNA owing to presence of the outrageous magnitude of mucilage and polysaccharides. Methods: A new fast track method for DNA isolation from the resting structure (sclerotia) of S. rolfsii by modifying CTAB method deprived of supplementation of proteinase K has been standardized. Result: The protocol produced 500ng DNA from sclerotia with purity vacillating from 1.7 to 1.9 as confirmed by A260/A280 and A260/A230 spectrophotometric documentation. The DNA extracted from sclerotia commissioning protocol was efficaciously used for the further downstream reactions/process like PCR-RAPD, PCR-ISSR and ITS amplification of rDNA-ITS region.