A rapid and efficient assay for extracting DNA from fungi.
OBJECTIVE
A method for the rapid extraction of fungal DNA from small quantities of tissue in a batch-processing format was investigated.


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.


CONCLUSIONS
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.
Griffin
D W
DW
United States Geological Survey, Center for Coastal and Regional Marine Studies, St Petersburg, FL 33701, USA. dgriffin@usgs.gov
Kellogg
C A
CA
Peak
K K
KK
Shinn
E A
EA
D004271:DNA, Fungal / Q000032:analysis*; D005615:Freezing; D005658:Fungi / Q000235:genetics*; D016133:Polymerase Chain Reaction
D004271:DNA, Fungal