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A rapid and efficient assay for extracting DNA from fungi
Author(s) -
Griffin D.W.,
Kellogg C.A.,
Peak K.K.,
Shinn E.A.
Publication year - 2002
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.2002.01071.x
Subject(s) - pipette , dna extraction , chromatography , dna , biology , extraction (chemistry) , plant tissue , boiling , polymerase chain reaction , microbiology and biotechnology , chemistry , botany , biochemistry , organic chemistry , gene
Aims: A method for the rapid extraction of fungal DNA from small quantities of tissue in a batch‐processing format was investigated. Methods and Results: Tissue (< 3·0 mg) was scraped from freshly‐grown fungal isolates. The tissue was suspended in buffer AP1 and subjected to seven rounds of freeze/thaw using a crushed dry ice/ethanol bath and a boiling water bath. After a 30 min boiling step, the tissue was quickly ground against the wall of the microfuge tube using a sterile pipette tip. The Qiagen DNeasy Plant Tissue Kit protocol was then used to purify the DNA for PCR/sequencing applications. Conclusions: The method allowed batch DNA extraction from multiple fungal isolates using a simple yet rapid and reliable assay. Significance and Impact of the Study: Use of this assay will allow researchers to obtain DNA from fungi quickly for use in molecular assays that previously required specialized instrumentation, was time‐consuming or was not conducive to batch processing.