1. Cut off 100 mg of the fresh plant tissue or 50 mg of the dry plant tissue.
2. Grind the sample in the liquid nitrogen to a fine powder using a mortar and pestle.
3. Add 400 μl of the Grind Buffer to the pestle and mortar and continue to homogenize the
sample tissue by grinding.
Step 1 Lysis
1. Transfer the mixture from the Grind Step to a 1.5 ml microcentrifuge tube.
2. Incubate at 70°C for 30 minutes to lyse the sample. During incubation, invert the tube
every 5 minutes.
3. Centrifuge for 5 minutes at 5,000 x g.
4. Transfer the supernatant to a new 1.5 ml microcentrifuge tube and add 200 μl of Lysis
Buffer. Mix well and incubate at 65°C for 5 minutes. During this time, pre-heat the
Release Buffer to 65ºC for Step 4.
5. Add 400 μl of the isopropanol to the lysate and mix well.
Step 2 DNA Binding
1. Add 20 μl of the 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 the magnetic beads to disperse completely during the binding, wash and elution