This protocol describes the isolation of genomic-DNA from 100 mg tissue.

Homogenization of sample:

Homogenize approx. 100 mg tissue to reduce to small pieces with a scalpel or better with a

commercial homogenizer.

1. Add 100 mg sample to a 1.5 mL microcentrifuge tube.

2. Add 1 mL Lysis & Binding Buffer, vortex for 30 seconds and incubate for 35 minutes at room

temperature.

Tip: Vortex the tube from time to time to get a complete lysis of the sample.

3. Spin for 5 minutes in a microcentrifuge at high speed (13,000 × g). Transfer the supernatant (liquid

phase) to a fresh 1.5 mL microcentrifuge tube.

Note: If floating material is present on top of the liquid, carefully pipet under it, avoiding aspiration of

floating material.

4. Add 100 µL MagIso™-DNA Particles to the supernatant, vortex and incubate for 5 minutes at room

temperature.

Tip: Before use resuspend the magnetic particles completely by vortexing

5. Place the tube on a magnetic separator for 30 seconds and collect the particle/DNA-pellet. Remove

and discard the supernatant.

Tip: Some of the solution with particles will end up in the cap of the tube. The tube can be tipped or

turned upside down, while placed in the magnet, to wash down the particles trapped in the cap.

6. Add 1 mL Wash Buffer I and vortex at room temperature. Collect the particle/DNApellet for 30

seconds with the magnet, remove and discard the supernatant. Repeat washing step once.

7. Add 1 mL Wash Buffer II and vortex for 5 seconds. Collect the particle/DNA-pellet for 30 seconds

with the magnet, remove and discard the supernatant. Repeat washing step once with Wash Buffer III.

Attention: During the wash process with Wash Buffer III vortex for 1 second.

8. Add 100 µL Elution Buffer (ddH2O), vortex and incubate for 10 minutes at 65 °C in a thermo-mixer

and vortex the tube from time to time for the complete resuspension of the pellet.

Tip: Complete resuspension of the pellet is important to recover high yields of DNA.

9. Collect the particles with the magnet and transfer the solution with the eluted DNA to a new clean

tube. If the solution is not clear, repeat the step to remove remaining magnetic particles.

Tip: The isolated DNA can be stored at 2-8 °C in a refrigerator, but for a long term storage - 20 °C is

recommended.

reference:

1.http://www.cd-bioparticles.com/p/Genomic-DNA-Purification-Kit_6_202_202_520.html

2.http://www.cd-bioparticles.com/pdf/WHK-C008.pdf

Is this kit your need? hope it can help~

Thanks Yudong, but I m looking for protocol for cultured cells, not tissue and not with magnetic beads, only columns. And for a small number of cells.

But thank you nevertheless for your answer.