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Viral gDNA Extraction Kit(SpinColumn)

Packing Specification：

Product Introduction：
Adopting spin columns that specifically bind viral DNA and a unique buffer system, this Viral DNA Extraction Kit is suitable for the rapid extraction of high-purity viral DNA from cell-free body fluids, including plasma, serum, ascites, cultured cell supernatant, cerebrospinal fluid, and urine. The product meets the extraction requirements of most viral DNAs, such as HBV (Hepatitis B Virus) and CMV (Cytomegalovirus).
After viral lysis, DNA is selectively adsorbed onto the silica membrane of the spin column under high chaotropic salt conditions. Through a series of rapid washing and centrifugation steps, impurities such as salts, cellular metabolites, and proteins are removed. Finally, pure viral DNA is eluted from the silica membrane using a low-salt Elution Buffer. The purified viral nucleic acid is free of impurities and PCR inhibitors, and can be directly used for PCR, enzyme digestion, hybridization, and other analyses.

Product Features：
1.The silica membrane in the spin column is made of high-quality special adsorption membrane, ensuring minimal variation in adsorption capacity between columns.
2.No need for toxic reagents such as phenol or ethanol precipitation steps.
3.Fast and simple; operation for a single sample can generally be completed within 20 minutes.
4.Multiple column washes ensure high purity. The extracted viral DNA has high purity and stable quality, suitable for various operations including PCR, enzyme digestion, and hybridization.

Application Scope：
Suitable for the rapid extraction of viral genomic DNA from cell-free body fluids, including plasma, serum, ascites, cultured cell supernatant, cerebrospinal fluid, and urine samples.

Operation Steps：
Tip: Before first use, add the specified amount of anhydrous ethanol to Wash Buffer WB, mix thoroughly, and mark the box to indicate ethanol has been added to avoid repeated addition!

1.Transfer 200 μL of serum or other body fluids (bring to room temperature first; make up to 200 μL with 0.9% NaCl or PBS if insufficient) into a 1.5 mL centrifuge tube. Add 400 μL of Lysis Buffer DLB and immediately vortex to mix thoroughly.

2.Incubate at room temperature (15-25℃) for 10 minutes, vortexing to mix once every 5 minutes.

3.Add 450 μL of anhydrous ethanol and immediately vortex to mix thoroughly.
If the ambient temperature exceeds 25℃, pre-cool the anhydrous ethanol on ice before adding.

4.Load the mixture into a Spin AC column (placed in a collection tube). Centrifuge at 13,000 rpm for 30-60 seconds and discard the waste liquid in the collection tube.
If the total volume exceeds 750 μL, load the solution into the same Spin AC column in two batches.

5.Add 500 μL of Deproteinization Buffer RE, centrifuge at 12,000 rpm for 30 seconds, and discard the waste liquid.

6.Add 500 μL of Wash Buffer WB (ensure anhydrous ethanol has been added!), centrifuge at 12,000 rpm for 30 seconds, discard the waste liquid, and repeat this step with another 500 μL of Wash Buffer WB.

7.Place the Spin AC column back into the empty collection tube and centrifuge at 13,000 rpm for 2 minutes to thoroughly remove residual Wash Buffer (residual ethanol may inhibit downstream reactions).

8.Remove the Spin AC column and place it into a new centrifuge tube. Add 30-50 μL of Elution Buffer EB (preheating in a 65-70℃ water bath beforehand improves elution efficiency) to the center of the adsorption membrane. Incubate at room temperature for 1 minute, then centrifuge at 12,000 rpm for 1 minute. For higher DNA yield, transfer the eluted solution back into the spin column and centrifuge at 12,000 rpm for 1 minute.
Larger elution volumes result in higher elution efficiency. For higher DNA concentration, reduce the elution volume (minimum 20 μL; volumes smaller than this reduce elution efficiency and DNA yield).

9.Store DNA at 2-8℃ for short-term use or -20℃ for long-term storage.