Data from: A crush on small fungi: an efficient and quick method for obtaining DNA from minute ascomycetes

1. We have developed a reliable technique for extracting DNA from single
microscopic fungal thalli, including efficient cell disruption and
transfer of cell content for subsequent polymerase chain reaction (PCR).
The technique was primarily developed for members of the ascomycete order
Laboulbeniales, which are minute fungi with tough cell walls that are
exceedingly difficult to disrupt with standard extraction techniques. 2.
Our method makes routine amplification of DNA from single thalli possible,
even from small species or poorly developed individuals. DNA release is
accomplished in an entirely mechanical manner using an arbor press fitted
with custom-made components. This approach has eliminated additional
treatment such as laborious freeze-thaw cycles, enzymes, or lysing agents.
3. The overall PCR success rate of 89% is comparable to or better than
alternative protocols that make use of substantially larger amounts of
fungal tissue. From 97% of the successful PCRs a total of 156 sequences
from four gene regions were produced. 4. Being able to restrict DNA
extractions to a single thallus is critical to all genetic studies
requiring data at the level of individual, e.g., population genetics.
Since all researchers working with minute uncultivable organisms in many
respects face the same problems (effective handling of the material, small
quantities of DNA etc.), the methodology described here has a potential to
be widely applicable. Necessary custom-made components can be manufactured
at fairly low cost by any precision-tool workshop using our detail
drawings.