OmniPrep™ for Fungus
To avoid repeated freezing‐thaw, dispense the Proteinase K solution into aliquots of 30µl/tube and freeze at ‐20°C.
If a precipitate forms due to cold storage allow to warm to room temperature until precipitate dissolves.
Centrifuge the Molecular Grinding Resin tube for 2 minutes at 2,500x g and remove the water.
Add 0.5ml Genomic Lysis Buffer.
Collect fungal tissue from liquid culture and wash 2‐3 times in sterile water.
Fungal mycelia are best prepared by grinding samples using Molecular GrindingResin™ in Genomic Lysis Buffer.
Add 10‐20mg fungal mycelia to a microcentrifuge tube containing 500µl Genomic Lysis Buffer.
Resuspend Molecular Grinding Resin by vigorous mixing or vortexing.
Add 30µl Molecular Grinding Resin™ using a wide bore pipette tips and grind with a microcentrifuge pestle.
Add 1µl Proteinase K solution for every 100µl Lysis Buffer and incubate at 60°C for 1‐2 hours.
Invert the tube periodically each hour.
Allow the sample to cool to room temperature.
Add 200µl chloroform and mix by inverting the tube several times.
Centrifuge for 10 minutes at 14,000xg and carefully remove the upper phase to a clean microcentrifuge tube.
Add 50µl DNA Stripping Solution to the sample and invert several times to mix.
Incubate the sample for 5‐10 minutes at 60°C.
Add 100µl Precipitation Solution and mix by inverting the tube several times.
A white precipitate should be produced, if not add 50µl aliquots of Precipitation Solution until a white precipitate forms.
Centrifuge the sample at 14,000xg for 5 minutes.
Transfer the supernatant to a clean tube and precipitate the genomic DNA with 500µl isopropanol.
Invert the tubes 10 times to precipitate the DNA.
Centrifuge at 14,000xg for 5 minutes to pellet genomic DNA.
Remove the supernatant.
Add 700µl 70% ethanol to the tube and invert several times to wash the DNA pellet.
Centrifuge for 1 minute at 14,000xg.
Decant or pipette off the ethanol wash.  
Invert the tube on a clean absorbent surface for several minutes to allow any excess ethanol to drain away.
Do not let the pellet dry completely or it will be difficult to rehydrate.
Add 50 to 100µl TE Buffer to the pellet.
Incubate at room temperature for at least 15 minutes to rehydrate.
Incubating the tube at 55‐60°C will speed up rehydration.
Incubate for 5‐60 minutes.
OPTIONAL: Add 1µl LongLife™ RNase for every 100µl TE Buffer at this stage.
Store DNA at 4°C, for long‐term storage store at ‐20°C or ‐80°C.
