Below you will find a simple and fast two-step DNA extraction protocol that works well with many different groups of fungi. This protocol can be utilized in preparation for DNA barcoding of fungi utilizing Oxford Nanopore MinION.
Varies depending on the use case. My samples generally have the metadata and images stored on iNaturalist. For each sample, I go to the iNaturalist observation, validate that the specimen matches the metadata and images, include a unique lab code in the "Voucher Number(s)" observational field, record the lab code on the sample packet with a marker, and record the lab code on the associated spreadsheet in the correct cell number.
If all of these actions need to be performed, it can take 3-5 hours per 96 samples to load.
If the iNaturalist observations have already been validated, the lab numbers have been assigned, and all that is required is to put a small amount of tissue in each cell for each sample, then the time outlay can be less than 2 hours per 96 samples.
There are a number of species groups that will have lower success rates with this protocol. They will either need a more complex DNA extraction protocol that involves grinding, or different primer combinations.
Low success rates (0-20% of specimens): Cantharellus, Helvella, Strobilomyces, Ophiocordyceps, Panellus
Moderate success rates (20-60% of specimens): Recalcitrant crusts & polypores, Cuphophyllus, Neohygrocybe, Tylopilus, Xylaria, Hypoxylaceae, Nidulariaceae
Overall, for a mix of general collections, we typically have success rates of 85-95% of specimens. The lowest success rate seen thus far for general collections was from MO-NAMA 2022, which had 75% success, despite being heavy on crust fungi and low-quality specimens.