Lichen Genomic DNA Extraction

Around one gram of collected target lichen samples from each specified sampling position mentioned above was weighed and rinsed by using an autoclaved Merck Millipore system (Burlington, MA, USA) produced high-purity water, Milli-Q ultrapure. After being cut into small fragments with sterilized scissors, the rinsed target lichen samples were subsequently pulverized using an autoclaved mortar and pestle. For DNA isolation, the Nippon Gene (Tokyo, Japan) produced ISOIL Large for Beads ver.2 device was employed for the isolation of bulk genomic DNA from the finely ground collected target lichen samples by using the bead-beating method, then add facilitator of precipitation, Ethachinmate (also produced by Nippon Gene) with 70% ethanol into the bulk DNA solution, following the past outlined procedure [12 (link)]. The resulting DNA precipitate was carefully redissolved in autoclaved Milli-Q ultrapure water. Purity and quantity assessments were performed by using the Thermo Fisher Scientific (Waltham, MA, USA) produced NanoDrop 2000c. Before the PCR amplification, the DNA samples were then securely stored at a temperature of − 20 °C.