A new simple method used to eliminate polysaccharides that cause problems during DNA isolation was established for 6 different white-rot fungi using 1% hexadecyltrimethylammonium bromide (CTAB) as wash buffer and followed by centrifugation. Variation in the DNA yield and quality was ascertained using precipitating agents, detergents and cell-wall-hydrolyzing chitinase. Considerable amount of exopolysaccharides from fungal biomass was removed with the use of 1% CTAB wash buffer followed by centrifugation. The DNA varied in terms of yield and quality. For the DNA extraction use of 2% SDS in extraction buffer worked best for Pycnoporus cinnabarinus, Cyathus bulleri, Cyathus striatus and Cyathus stercoreus, while 2% CTAB worked best for Phanerochaete chrysosporium and Pleurotus ostreatus. Elimination of phenol and use of absolute ethanol for precipitating DNA resulted in good yield and quality of DNA. This DNA was amenable to restriction endonuclease digestion.

Department of Microbiology University of Delhi South Campus New Delhi 110 021 India

See more in PubMed

Folia Microbiol (Praha). 2002;47(6):691-5 PubMed

Folia Microbiol (Praha). 2002;47(4):417-21 PubMed

Nucleic Acids Res. 1980 Oct 10;8(19):4321-5 PubMed

Biotechniques. 1997 Jan;22(1):47, 50-2 PubMed

J Gen Microbiol. 1978 Jun;106(2):387-9 PubMed

Folia Microbiol (Praha). 2002;47(4):385-90 PubMed

Biotechniques. 1991 Feb;10(2):162, 164, 166 PubMed

FEMS Microbiol Lett. 1990 Jan 1;54(1-3):113-7 PubMed

Anal Biochem. 1983 Dec;135(2):416-22 PubMed

Prep Biochem. 1995 Nov;25(4):171-81 PubMed

Anal Biochem. 1982 Jan 1;119(1):158-63 PubMed

Folia Microbiol (Praha). 2003;48(3):413-6 PubMed

Gene. 1997 Sep 1;196(1-2):113-9 PubMed

Annu Rev Microbiol. 1987;41:465-505 PubMed

Adv Biochem Eng Biotechnol. 1997;57:45-125 PubMed

Appl Environ Microbiol. 1999 Nov;65(11):4943-8 PubMed

A simple and efficient genetic transformation method of Ganoderma weberianum

Improved method of detection and molecular typing of Borrelia burgdorferi sensu lato in clinical samples by polymerase chain reaction without DNA purification