Tissue Genomic DNA Kit Protocol
1. Cut off up to 30 mg of the animal tissue and transfer it to a 1.5 ml microcentrifuge tube.
2. Add 200 μl of the Grind Buffer to the tube and continue to homogenize the sample tissue by grinding.
Step 1 Lysis
1. Add 20 μl of the Proteinase K (10 mg/ml) to the sample mixture and mix by vortex.
2. Incubate at 65°C for 30 minutes to lyse the sample. During the 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 300 μl of the Lysis Buffer.
5. Mix well and incubate at 65°C for 5 minutes. During this time, pre-heat the Release Buffer to 65°C for the Step 4.
6. Add 300 μl of the absolute EtOH 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 the magnetic bead separation time).
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 65°C) and mix well.
2. Incubate for 3 minutes at 65°C (During the incubation, shake the tube vigorously every minute).
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 steps.