Virus Nucleic Acid Isolation Kit

The Virus Nucleic Acid Isolation Kit provides a fast, simple, and cost-effective method for the isolation of viral DNA/RNA from cell-free samples such as serum, plasma, body fluids and the supernatant of virus-infected cell cultures. Its unique buffer system will efficiently lyse cells and degrade protein, allowing for the nucleic acid to be easily bound by the glass fiber matrix of the column. Contaminants such as salts, metabolites and soluble macromolecular cellular components are removed in the Wash Step. The phenol extraction and ethanol precipitation are not required, and the high-quality nucleic acid is eluted in the RNase-free elution buffer. The viral DNA/RNA isolated with the Total Nucleic Acid Isolation Kit (Virus) is suitable for a variety of routine applications, including the Real-time PCR/RT-PCR, Automated Fluorescent DNA Sequencing, PCR, and other enzymatic reactions. The entire procedure can be completed within 15-20 minutes.

Kit Contents

*It is a normal condition if Buffer V1 has found visually turned into yellow, which does not affect the function of this product

• Isolates both viral RNA and DNA, allowing simultaneous detection of both types of virus.
• Removes inhibitors that might interfere with downstream assays, ensuring greater assay specificity, sensitivity and reproducibility.
• Prepares nucleic acid samples in only 20 minutes.
• Yields a concentrated sample that is suitable for direct assay (no precipitation required).
• Universal viral nucleic acid purification system: One kit for both DNA and RNA viral purification, allowing simultaneous testing of both viral types.
• Environment-friendly: Less infectious plastic waste due to the reduced number of hands-on steps.
• Safety: No phenol/chloroform extractions.
• Versatility: Spin and vacuum formats available.
• Quality: RNA suitable for downstream applications.

• Add 45 ml of the ethanol (96-100%) to the Buffer V2 and shake before use.
• Add 60 ml of the ethanol (96-100%) to the Buffer W2 and shake before use.

Step 1 Lysis

1. Transfer up to 200 μl of the virus sample into a 1.5 ml microcentrifuge tube and add 400 μl of the Buffer V1. (If the sample is less than 200 μl, adjust the sample volume to 200 μl with the PBS)
2. Mix well and let it stand at the room temperature for 10 minutes.

Step 2 Nucleic Acid Binding

1. Add 450 µl of the Buffer V2 (ethanol added) to the sample lysate and shake vigorously.
2. Place a Column VN in a 2 ml Collection Tube.
3. Transfer 700 µl of the lysate mixture into the Column VN.
4. Centrifuge at 16,000 x g for 1 minute.
5. Discard the flow-through and place the Column VN back in the same Collection Tube.
6. Transfer the remaining lysate mixture to the Column VN.
7. Centrifuge at 16,000 x g for 1 minute.
8. Discard the flow-through and place the Column VN back in the same Collection Tube.

Step 3 Wash

1. Add 400 µl of the Buffer W1 into the Column VN.
2. Centrifuge at 16,000 x g for 30 seconds.
3. Discard the flow-through and place the Column VN back into the same Collection tube.
4. Add 600 µl of Buffer W2 (ethanol added) into the Column VN.
5. Centrifuge at 16,000 x g for 30 seconds.
6. Discard the flow-through and place the Column VN back into the same Collection tube.
7. Centrifuge at 16,000 x g again for 2 minutes to remove the residual Buffer W2.

Step 4 Elution

1. Place the Column VN in a clean 1.5 ml microcentrifuge tube (DNase and RNase free).
2. Add 50 μl Buffer RE or RNase-free water (pH is between 7.0 and 8.5) to the center of each Column VN, let it stand for 2 minutes, and centrifuge at 14,000 x g for 2 minutes.

• It is a normal condition if Buffer V1 has found visually turned into yellow, which do not affect the function of this product.
• Check the Buffers before use for salt precipitation. Re-dissolve any precipitate by warming up to 37°C.
• The Buffers V1 and W1 contain irritants. Wear gloves when handling these buffers.