Mag-Bind Soil DNA Kit Protoco

1. Weigh 500 mg of glass beads in a 2 ml centrifuge tube, add 0.2-0.5 g soil sample. Add 0.8 ml Buffer SLX Mlus. Vortex at maximum speed for 3-5 minute to lyse samples. For the best result, A Mixer Mill, such as Fastprep-24, Mixer Mill MM 300, should be used.   2. Add 80 ul Buffer DS and vortex to mix.   3. Incubate at 70°C for 10 min. Briefly vortex the tube once during the incubation. For some difficult lysis bacterial, Increase the temperature to 90°C.   4. Centrifuge at 13,000 x g for 5 min. Transfer 600 μL the supernatant into a new 2 ml tube and add 200 μL Buffer SP2. Mix the sample throughly by vortexing.   5. Incubate on ice for 5 minutes. Centrifuge at 13,000 x g in a microcentrifuge for 5 minutes.   6. Carefully transfer supernatant to a new 2 ml tube and add100 ul HTR Reagent. Mix throughly by vortexing for 10 seconds.   7. Incubate at room temperature for 2 minutes. Centrifuge at full speed (13,000 x g) for 2 minutes.   8. Transfer 500 ul of the cleared supernatant to a new 1.5 ml tube.   9. Add 500 ul Binding Buffer and 60 ul of MagSi Particles to the sample. Invert to mix well. Incubate at room temperature for 2 minutes.   10. Place the tube on a magnetic separation device suitable for 2 mL tube to magnetize the MagSi particles. Carefully remove and discard the cleared supernatant.   11. Remove the tube containing the MagSi? particles from the magnetic separation device.   12. Add 1000 ul Buffer PHB into the tube. Resuspend MagSi particles pellet by vortexing.   13. Place the tube onto a magnetic separation device to magnetize the MagSi particles. Carefully remove and discard the cleared supernatant.   14. Add 1000 ul of SPM Wash Buffer diluted with ethanol into the tube. Resuspend Mag-Bind particles pellet by vortexing.   15. Place the tube onto a magnetic separation device to magnetize the MagSi particles. Carefully remove and discard the cleared supernatant.   16. Wash MagSi particles with SPM one more time by repeating step 14-15.   17. After remove the supernatant, air dry the magnetic beads by invert the tube on a absorbent paper for 15 minutes. Remove any residue liquid from tube with pipettor.   18. Add 50-100 ul Elution Buffer or water to the tube. Incubate the tube at Resuspend MagSi  particles by vortexing. Incubate at 65°C for 10 minutes if maxium DNA yield is desired.   19. Place the tube onto a magnetic separation device to magnetize the Mag-Bind? particles.   20. Transfer the cleared supernatant containing purified DNA to a new 1.5 mL tube.