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A Fast and Inexpensive High-Throughput Protocol for Isolating High Molecular Weight Genomic DNA from Lichens

Author:
Werth, Silke, Reynisdóttir, Sigrún, Gudmundsson, Hörður, Andrésson, Ólafur S.
Source:
Herzogia 2016 v.29 no.2 pp. 610-616
ISSN:
0018-0971
Subject:
Ascomycota, Basidiomycota, DNA, centrifugation, cetyltrimethylammonium bromide, ethanol, fungi, genomic libraries, glass fibers, guanidines, lichens, molecular weight
Abstract:
Isolating high molecular weight DNA as required for genomic library preparations and other applications is a challenging task. We optimized a glass-fiber column-based, high throughput DNA isolation protocol to yield comparatively large quantities of high molecular weight, pure DNA; cell lysis based on a CTAB buffer was followed by centrifugation to remove cell debris, binding to a glass-fiber membrane under high concentration of a chaotropic salt (guanidine hydrochloride), followed by two 70 % ethanol washes and elution. This method was utilized successfully on several species of lichenized ascomycetes and on two non-lichenized basidiomycetes. Thus, while developed for lichens, the method is suitable for a range of fungal taxa.
Agid:
5600652