DNA Extraction from Fungal Organisms

Instrument: Minilys Personal Homogenizer

Sample: Coprinopsis cinerea and Talaromyces emersonii

Protocol:

C. cinerea and T. emersonii were grown in both solid and liquid state cultures following standard protocols (Binninger et al., 1987; Gupta et al., 2012). 0.5g of mycelia scrapped from solid culture and 0.4g of mycelia harvested from liquid culture were used for DNA extractions.
Mycelia samples were placed into the Precellys® vial containing ceramic/glass beads, using a sterile pipette tip to help push mycelia into contact with the tubes contents.
Samples were homogenize for 60 seconds in the Minilys® bead mill homogenizer at the low/medium setting until the contents were seen to be blitzed.
600 µL SFG1 Buffer and 4 µL RNase A (E.N.Z.A SP Fungal DNA Mini kit D5542-01) were added to the vial, and vortexed to ensure that samples were suspended and that no clumps remained.
The extraction was carried out from step 3 of the E.N.Z.A SP Fungal DNA Mini kit, according to manufactures instructions.

Results:

DNA extractions were performed on both T. emersonii and C. cinerea mycelia harvested from liquid culture. The combination of the Minilys® personal homogenizer with the E.N.Z.A fungal DNA extraction kit resulted in a successful and rapid DNA
extraction from both organisms. In order to increase efficiency for high throughput screening, the method was then tested on mycelia scraped from solid state cultures. DNA was successfully extracted from both organisms thus eliminating the need for liquid cultivation.

Figure 1: DNA extraction from plate mycelia performed using Minilys® bead mill, Precellys® lysing kit tubes and E.N.Z.A. fungal extraction kit on Coprinopsis cinerea FA2222. 1% agarose gel with gel red, lane 1: C. cinerea DNA, lane 2&3: 1kb molecular weight ladder.

Institute of Technology, Sligo, Ireland