About Quick-DNA™ Viral 96 Kit
The Quick-DNA™ Viral 96 Kit provides for the rapid isolation of high-quality viral DNA from a wide range of biological sources. A uniquely designed buffer is included for the efficient denaturation of viral particles in whole blood (fresh and stored), plasma, serum, tissue, ascites, cultured cells, and from liquid samples. DNA can be eluted with elution buffer or water and is suitable for subsequent PCR, nucleotide blotting, and restriction endonuclease digestion procedures.
|Format||96-well Spin Plate|
|DNA Recovery||Typically, up to 5 μg total DNA (per well) is eluted into 10-15 μl DNA Elution Buffer. For DNA 75 bp to 10 kb, the recovery is 70-90%. For DNA 11 kb to 50 kb the recovery is 50-70%.|
|Processing Time||25 min|
|Equipment||Centrifuge with microplate carriers|
|DNA Size Limits||100 bp to 50 kb|
|Sample Sources||Serum, plasma, tissue, or cells derived from humans, mice, rabbits, etc.|
|DNA Purity||High quality DNA is eluted with DNA Elution Buffer making it perfect for PCR. A260/A280 >1.8|
Viral DNA purification. Human HBV DNA was isolated from 10 to 0.001 µl of human serum using phenol/chloroform or Quick-DNA™ Viral 96 Kit. The presence of HBV DNA is evidenced by a ~200 bp PCR amplicon. Lane M is a 100 bp DNA Ladder and "Neg." is the negative control for PCR.
Add 4 volumes of ZR Viral DNA Buffer to each volume of sample (e.g., 800 µl ZR Viral DNA Buffer to 200 µl serum). Allow to stand at room temperature for 5-10 minutes.
It may be necessary to centrifuge the sample/ZR Viral DNA Buffer mixture before transferring the supernatant to a well of the Zymo-Spin™ I-96 Plate to remove particulate matter that may clog the matrix.
Longer incubation periods may be required for some samples.
Transfer sample mixtures to the wells of a Zymo-Spin™ I-96 Plate mounted on a Collection Plate. Centrifuge at ≥2,500 x g (5,000 x g max.) for 5 minutes until sample mixtures have been completely filtered. Discard the flow through.
Add 300 µl DNA Wash Buffer to each well of the Zymo-Spin™ I-96 Plate. Centrifuge at ≥2,500 x g for 5 minutes. Repeat wash step.
Add 10-15 µl Elution Buffer or water directly to the column matrix in each well. Transfer the Zymo-Spin™ I-96 Plate onto a DNA Elution Plate and centrifuge at ≥2,500 x g for 5 minutes to elute the DNA.
Elution of DNA from the column is dependent on pH and temperature. If water is used, ensure the pH is <6.0. Also, the total yield may be improved by eluting the DNA with Elution Buffer or water pre-equilibrated to 60-70oC.
Alternatively, for 96-well PCR, viral DNA can be eluted with water directly from the Zymo-Spin™ I-96 Plate into a 96-well PCR plate. Simply, mount your 96-well PCR plate between the Zymo-Spin™ I-96 Plate and an empty Collection Plate then elute the DNA as described above.
DNA is now ready for immediate use or storage.