Open AccessRediscovering an old foe: Optimised molecular methods for DNA extraction and sequencing applications for fungarium specimens of powdery mildew (Erysiphales)
Author(s) -
Rean L. Smith,
Tim Sawbridge,
Ross Mann,
Jatinder Kaur,
Tom W. May,
Jacqueline Edwards
Publication year - 2020
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0232535
Subject(s) - powdery mildew , dna extraction , biology , dna , phylogenetic tree , dna sequencing , extraction (chemistry) , ascomycota , computational biology , botany , genetics , chromatography , chemistry , polymerase chain reaction , gene
The purpose of this study was to identify a reliable DNA extraction protocol to use on 25-year-old powdery mildew specimens from the reference collection VPRI in order to produce high quality sequences suitable to address taxonomic phylogenetic questions. We tested 13 extraction protocols and two library preparation kits and found the combination of the E.Z.N.A. ® Forensic DNA kit for DNA extraction and the NuGen Ovation ® Ultralow System library preparation kit was the most suitable for this purpose.