Mbead Bacteria Genomic DNA Kit

The MBead Bacteria Genomic DNA Kit provides a fast, simple, and cost-effective method for isolating the genomic DNA from bacterial cells. Its unique buffer system will efficiently lyse cells and degrade protein, allowing for DNA to be easily bound by the surface of the magnetic beads. The Phenol extraction and ethanol precipitation are not required, and the high-quality genomic DNA is eluted in the Release Buffer. The genomic DNA purified with the MBead Bacteria Genomic DNA Kit is suitable for a variety of routine applications. The entire procedure can be completed within 15-20 minutes (using most magnetic separators) or the kit can be easily adapted to satisfy most automated Nucleic Acid purification systems.

• Sample: Up to 300 µl of bacteria.
• Applications: Restriction Enzyme Digestion, Southern Blotting, PCR and qPCR assays.
• Operation time: within 15 minutes (manual).
• Storage: room temperature.

Kit Contents

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 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 65ºC) and mix well.
2. Incubate for 3 minutes at 65ºC (during 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 Magnetic Beads to disperse completely during the binding, wash and elution steps.

• Absolute EtOH
• Magnetic separator
• 1.5 ml microcentrifuge tubes
• Water bath / Dry bath

• During operation, always wear a lab coat, disposable gloves, and protective equipment.
• Research Use Only. Not intended for any animal or human therapeutic or diagnostic uses.