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Rapid and reliable high‐throughput methods of DNA extraction for use in barcoding and molecular systematics of mushrooms
Author(s) -
DENTINGER BRYN T. M.,
MARGARITESCU SIMONA,
MONCALVO JEANMARC
Publication year - 2010
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.1755-0998.2009.02825.x
Subject(s) - biology , dna barcoding , systematics , dna extraction , evolutionary biology , extraction (chemistry) , computational biology , zoology , genetics , taxonomy (biology) , gene , polymerase chain reaction , chromatography , chemistry
We present two methods for DNA extraction from fresh and dried mushrooms that are adaptable to high‐throughput sequencing initiatives, such as DNA barcoding. Our results show that these protocols yield ∼85% sequencing success from recently collected materials. Tests with both recent (<2 year) and older (>100 years) specimens reveal that older collections have low success rates and may be an inefficient resource for populating a barcode database. However, our method of extracting DNA from herbarium samples using small amount of tissue is reliable and could be used for important historical specimens. The application of these protocols greatly reduces time, and therefore cost, of generating DNA sequences from mushrooms and other fungi vs. traditional extraction methods. The efficiency of these methods illustrates that standardization and streamlining of sample processing should be shifted from the laboratory to the field.