Mbead Plant Genomic DNA Kit

This MBead Plant Genomic DNA Kit was designed specifically for genomic DNA isolation from a variety of plant samples. 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 RNA and other non-specific binding particles are removed with a wash buffer, and the genomic DNA is then released into the Release Buffer. The genomic DNA can be purified manually within 50 minutes (using most magnetic separators) or the kit can be easily adapted to satisfy most automated Nucleic Acid purification systems.

• Sample: Up to 50 mg of various plant tissue
• Applications: Restriction Enzyme Digestion, Southern Blotting, PCR, qPCR and RT-PCR assays
• Operation time: Within 50 minutes (manual)
• Storage: Room temperature

Kit Contents

Sample Preparation

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 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.

• Mortar and pestle
• Liquid nitrogen
• Isopropanol
• Magnetic separator
• 1.5 ml microcentrifuge tubes

• Check buffers before use for salt precipitation. Re-dissolve any precipitate by warming up to 37°C
• 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.