Step 1 Lysis
1. Transfer up to 300 μl of the bacterial culture into a 1.5 ml microcentrifuge tube and add
300 μl of Lysis Buffer.
2. Mix well and incubate at 65°C for 5 minutes. During this time, pre-heat the Release
Buffer to 65ºC for Step 4.
3. Add 300 μl of absolute EtOH to the lysate and mix well.
Step 2 DNA Binding
1. Add 20 μl of Magnetic Beads. Mix well by gently shaking for 3 minutes.
2. Place the tube in a magnetic separator for 30 seconds.
3. Remove the solution (if the mixture becomes viscous, increase magnetic bead separation
Step 3 Wash
1. Add 800 μl of the Wash Buffer and mix well (following the wash, the mixture will no
longer be viscous).
2. Place the tube in a magnetic separator for 30 seconds. Remove the solution.
Step 4 Release
1. Add 200 μl of the Release Buffer (pre-heated to 65ºC) and mix well.
2. Incubate for 3 minutes at 65ºC (during incubation, shake the tube vigorously every
3. Place the tube in a magnetic separator for 1 minute.
4. Carefully transfer ONLY the clean portion of the solution to a clean tube.
NOTE: Be sure and allow Magnetic Beads to disperse completely during the binding, wash and elution steps.