Mbead Buffy Coat Genomic DNA Kit

MBead Buffy Coat Genomic DNA Kit was designed specifically for genomic DNA isolation from Buffy Coat samples. Its special buffer system will efficiently lyse cell and degrade protein, allowing for DNA to be easily bound by the surface of the magnetic beads. The other non-specific binding particles are removed with a wash buffer, and the genomic DNA is released from the beads by addition of a low ionic strength buffer and heat. Genomic DNA can be purified manually within 15 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 the buffy coat.
• Applications: restriction Enzyme Digestion, Southern Blotting, PCR and qPCR assays.
• Operation time: 10-15 minutes (manual).
• Storage: room temperature.

Kit Contents

Step 1 Lysis

1. Transfer up to 300 μl of the Buffy Coat into a 1.5 ml microcentrifuge tube and add 300 μl of the Lysis Buffer.
2. 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.
3. Add 300 μl of the 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 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.

Refer to the table below to troubleshoot problems that you may encounter when purifying the genomic DNA with the kit.

• 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.