Yeast DNA extraction using DNeasy® Blood & Tissue Kit
Centrifuge a maximum of cells for 5 minutes at 300g (190rpm). Resuspend in 200µL PBS. Add 20µL zymolyase.
Add 200µL Buffer AL. Mix thoroughly by vortexing.
Add 200µL ethanol (96%). Mix thoroughly by vortexing.
Pipet the mixture into a DNeasy Mini spin column placed in a 2mL collection tube. Centrifuge at 6000g (8000rpm) for 1 minute. Discard the flow-through and collection tube.
Place the spin column in a new 2mL collection tube. Add 500µL Buffer AW1. Centrifuge for 1 minute at 6000g. Discard the flow-through and collection tube.
Place the spin column in a new 2mL collection tube. Add 500µL Buffer AW2. Centrifuge for 3 minutes at 20,000g. Discard the flow-through and collection tube.
Transfer the spin column to a new 1.5mL or 2mL microcentrifuge tube.
Elute the DNA by adding 200µL Buffer AE or sterile water to the center of the spin column membrane. Incubate for 1 minute at room temperature. Centrifuge for 1 minute at 6000g.
