About Quick-DNA™ Viral Kit
The Quick-DNA™ Viral 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.
|DNA Recovery||Typically, up to 5 μg total DNA is eluted into 6-10 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||15 min|
|DNA Size Limits||100 bp to 50 kb.|
|Sample Sources||Whole blood, 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 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.
In a 1.5 ml microcentrifuge tube, 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). Mix briefly by vortexing. Allow to stand at room temperature for 5-10 minutes.
Longer incubation periods may be required for some samples.
Transfer the mixture to a Zymo-Spin™ IC Column in a Collection Tube. Centrifuge for 1 minute. Discard the flow through from the Collection Tube.
It may be necessary to centrifuge the sample/ZR Viral DNA Buffer mixture before transferring the supernatant to the Zymo-Spin™ Column to remove particulate matter that may clog the column.
Add 300 µl DNA Wash Buffer to the column. Centrifuge for 1 minute. Discard the flow through. Repeat this step.
Place the Zymo-Spin™ IC Column into a microcentrifuge tube. Add 6-10 µl DNA Elution Buffer or water directly to the column matrix. Let stand at room temperature for 1 minute. Centrifuge for 1 minute 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 higher volumes (20-30 µl) Elution Buffer or water pre-equilibrated to 60-70oC.
DNA is now ready for immediate use or storage.