I have always used the DNeasy Plant Mini Kit for extracting fungal DNA. This is a rather long protocol and did not seem to work for one of my cultures. I was wondering if anyone has used Lyse and Go on their fungal cultures? I wanted to give it a shot. According to the protocol, it eliminates the lengthy DNA protocols and excessive sample manipulations.
It might work without that reagent as well, depending on cultures. Simply dip a toothpick on the edge of a culture (young mycelium) to take up some mycelium and then swirl into the pcr-reaction mix (should contain BSA)
I tried it for two different fungi but results were not consistent, sometimes it worked sometimes didn't.
I am using a simple protocol:cut a small piece of fungal agar culture (5 mm in diameter on PDA or CMA 1/10 strength), place it in a vial with 0.5 ml TE (10 mM Tris, 1 mM Na2EDTA, pH 8.0) and approx. 0.25 ml of 0.5 mm ceramic beads and heat at 95 C for 10 min. Then homogenize for 30s at full speed on vortex or in Fastprep 24 if available. Centrifuge and use supernatant undiluted or 10-100 x diluted as PCR template. Works perfectly for all kind of fungi.
Thank you for the help! I will be certainly be trying some of these suggestions.
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