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A simple method for extraction of fungal genomic DNA
Author(s) -
AlSamarrai T. H.,
Schmid J.
Publication year - 2000
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1046/j.1472-765x.2000.00664.x
Subject(s) - ethanol precipitation , mycelium , biology , chromatography , aspergillus niger , aspergillus flavus , dna extraction , polysaccharide , agar , chloroform , botany , microbiology and biotechnology , food science , chemistry , biochemistry , polymerase chain reaction , bacteria , genetics , gene
T. H. AL‐SAMARRAI and J. SCHMID.2000.We have developed a new, simple and effective method for extraction of fungal genomic DNA. The initial steps involved suspension of freeze‐dried mycelium in buffer containing sodium dodecyl sulphate, detachment of DNA from polysaccharides by mild shearing, NaCl precipitation of polysaccharides and protein, chloroform extraction and ethanol precipitation. The ethanol precipitate was then subjected to a second round of mild shearing, NaCl precipitation, chloroform extraction and ethanol precipitation. The procedure required approximately 1 h to perform. The method yielded 8–32 μg of high molecular weight DNA per 30 mg of freeze‐dried mycelium when tested on six fungal species: Aspergillus niger, A. flavus, Fusarium graminarum, Neotyphodium lolii, Penicillium citrinum and Rhizopus nigricanes. The DNA was digestible with Eco RI, Hind III, Sal I and Bam HI. For the slow‐growing N. lolii, a modification of the method was developed that removed the agar residue from colonies grown on agar plates by centrifugation at 13 000 rev min −1 in the presence of CsCl. The modified method yielded 1·5–2 μg of high molecular weight DNA per colony.