DNeasy Plant Mini Kit (250)

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	Follow manufacturer's instructions

Protocol tips
Follow manufacturer's instructions

Publication protocol

Freeze-thaw extraction of fungal DNA: Isolates from -70°C storage were streaked out onto plates of R2A agar and grown for 2 days at room temperature. Approximately 2 - 5 mg of fungus tissue from each isolate (see Fig. 1) were placed in a sterile 1-5 ml microcentrifuge tube. To each tube, 400 ll AP1 buffer (DNeasy Plant Mini Kit, Qiagen) and 4 ll RNase A (supplied with the kit) were added. Freeze/thaw was used to lyse fungal cells. This process was repeated seven times using crushed dry ice/ ethanol and a boiling water bath. After the last thaw step, the samples were boiled for 30 min. A sterile 1 ml micropipette tip was then used to grind any visible tissue in the tubes briefly (5 s) between the tip and conical bottom of the microcentrifuge tube. The DNeasy Plant Mini Kit ‘Protocol for Isolation of DNA from Plant Tissue with the DNeasy Plant Mini Kit’ procedure starting with Step 4 (add 130 ll of Buffer AP2...) was then followed. The DNA was eluted in 50 ll buffer AE and 5 ll of eluted DNA was used for PCR.

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Manufacturer protocol

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