About ZR Viral DNA/RNA Kit™
The ZR Viral DNA/RNA Kit™ provide for rapid, single column isolation of high-quality viral nucleic acids from a wide range of biological sources. The kit can be used to successfully isolate viral DNA and RNA from cell-free body fluids as well as cellular suspensions at concentrations ≤ 1 x 105 cells/ml. The procedure employs a single buffer system that facilitates viral particle lysis and allows for the subsequent DNA/RNA binding onto the matrix of the Zymo-Spin™ IC Column. The nucleic acids are washed then eluted with DNase/RNase-free Water. The eluted DNA and RNA are suitable for use in various subsequent procedures including RT/PCR.
| Format | Spin Column |
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| DNA Recovery | Up to 10 μg total DNA and/or RNA per column can be eluted with ≥ 10 µl water. |
| Processing Time | 5 min |
| Equipment | Microcentrifuge |
| DNA Size Limits | Capable of purifying small DNA/RNA fragments > 50 bases and large sized DNA/RNA > 200 kb. |
| Sample Sources | Cells, serum, plasma, culture supernatants. |
| DNA Purity | RT/PCR quality DNA and RNA (A260/A280 > 1.8, A260/A230 > 1.8) is recovered. |
| Storage | Eluted DNA/RNA can be stored at ≤ -70°C |
| RNA Purity | RT/PCR quality DNA and RNA (A260/A280 > 1.8, A260/A230 > 1.8) is recovered. |
| RNA Recovery | Up to 10 μg total DNA and/or RNA per column can be eluted with ≥ 10 µl water. |
| Sample Size | ≤ 200 µl (e.g. plasma, serum) can be processed per standard preparation. |
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Viral nucleic acids were isolated from liquid samples using the ZR Viral DNA/RNA Kit™. Isolated viral nucleic acids were reverse transcribed/amplified using a coupled RT-real-time PCR system (Zymo Research). Ct values for measles, influenza type A (FluA), and herpes-simplex (HSV) viruses were 23.05 (diamonds), 24.56 (triangles), 22.92 (circles), respectively. Negative control - RT-PCR (no template w/ HSV specific primers). Positive control - PCR (HSV template w/ HSV primers).
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| Step 1 | Add 3 volumes of Viral DNA/RNA Buffer to each volume of sample (e.g., 300 µl lysis buffer to 100 µl plasma or serum). |
|---|---|
| Step 2 | Transfer the mixture to the Zymo-Spin™ IC-XL Column1,2 in a Collection Tube and centrifuge at ≥12,000 x g for 1 minute. Discard the Collection Tube containing the flow-through. Place the column into a new Collection Tube. |
| Step 3 | Add 400 µl DNA/RNA Prep Buffer to the column and centrifuge at ≥12,000 x g for 30 seconds. Discard the flow-through. |
| Step 4 | Add 700 µl DNA/RNA Wash Buffer to the column and centrifuge at ≥12,000 × g for 15 seconds. Discard the flow-through. |
| Step 5 | Repeat Step 4 with 400 µl DNA/RNA Wash Buffer. Discard the flow-through. |
| Step 6 | Centrifuge Zymo-Spin™ IC-XL Column at maximum speed for 5 minutes in the emptied Collection Tube to ensure complete removal of the wash buffer. Remove the column carefully from the Collection Tube and transfer it into a DNase/RNase-Free Tube. |
| Step 7 | Add ≥10 µl DNase/RNase-Free Water3 directly to the column matrix and let stand for 1 minute at room temperature. Centrifuge at maximum speed for 30 seconds to elute DNA/RNA. |
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“The ZR-96 viral RNA kit performed much better than the RNeasy 96 kit for extracting RNA from dilute samples of virus supernatant. The kit is designed for small-volume elution to automatically concentrate the sample, and it’s designed for viral supernatants rather than cell lysates, so we didn’t need to deal with optimizing added carrier RNA. So in all, it was simpler, easier, and gave us a 10-fold improvement in our limit of detection.”
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S.B., Boehringer Ingelheim, Canada
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