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An inexpensive high‐throughput method to extract high yields of good quality DNA from fungi
Author(s) -
KIKUCHI T.,
KARIM N.,
MASUYA H.,
OTA Y.,
KUBONO T.
Publication year - 2009
Publication title -
molecular ecology resources
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.96
H-Index - 136
eISSN - 1755-0998
pISSN - 1755-098X
DOI - 10.1111/j.1755-0998.2008.02184.x
Subject(s) - biology , dna extraction , polymerase chain reaction , throughput , genomic dna , restriction enzyme , membrane filter , dna , extraction (chemistry) , population , chromatography , membrane , gene , biochemistry , computer science , telecommunications , chemistry , demography , sociology , wireless
We developed an efficient method for high‐throughput extraction of high‐quality DNA from various fungi. In this method, fungal mycelia were cultured and harvested on the surfaces of membranes on media plates. We degraded cell walls using a lytic enzyme (Yatalase). Purification was performed on 96‐well glass fibre filter plates. DNA was successfully extracted from various fungi provided (102 genus 132 species) at high yields and quality, and proved suitable for storage, polymerase chain reaction amplification and restriction enzyme digestion. The method described is rapid, inexpensive and automation friendly. This enables the simultaneous extraction of large numbers of samples, significantly improving the potential throughput in genomics, particularly in diagnostic and population studies.