Viral DNA/RNA Extraction Kit(SpinColumn)
Packing Specification：
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R2124-50
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Viral DNA/RNA Extraction Kit(SpinColumn)
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50T
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CNY1190
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R2124-200
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Viral DNA/RNA Extraction Kit(SpinColumn)
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200T
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CNY4280
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For research use only. Not for use in medicine, clinical diagnosis, food, cosmetics or other applications.
Product Introduction:
Using spin adsorption columns that specifically bind to viral DNA/RNA and a unique buffer system, the Viral DNA/RNA Extraction Kit is suitable for rapidly extracting high-purity viral DNA/RNA from cell-free body fluids, including plasma, serum, ascites, cultured cell supernatant, cerebrospinal fluid, and urine. This product can meet the requirement of simultaneous extraction of most viral RNA/DNA, such as viral RNA (HCV -Hepatitis C Virus, HIV-Human Immunodeficiency Virus, and HTLV-Human T-Lymphotropic Virus) and viral DNA (HBV-Hepatitis B Virus and CMV-Cytomegalovirus). After viral lysis, DNA/RNA selectively binds to the silica matrix membrane inside the spin column under high - chaotropic salt conditions (Poly Carrier is specially provided to easily capture trace nucleic acids from the system). Then, through a series of rapid washing - centrifugation steps, impurities such as salts, cellular metabolites, and proteins are removed. Finally, the pure viral DNA/RNA is eluted from the silica matrix membrane using a low - salt elution buffer. The purified viral nucleic acids are free of impurities and PCR inhibitors and can be directly used for PCR/RT - PCR analysis.
Product Features:
1.The silica matrix membranes inside the spin adsorption columns are all high - quality custom - made adsorption membranes, with minimal differences in adsorption capacity between columns.
2.There is no need to use toxic reagents such as phenol or perform steps like ethanol precipitation.
3.It saves time and is simple to operate; the operation for a single sample can generally be completed within 20 minutes.
4.Multiple column washing steps ensure high purity. The extracted viral DNA/RNA has high purity and stable and reliable quality, and can be used for various routine operations, including PCR/RT-PCR, enzyme digestion, sequencing, and Southern hybridization.
Operation Steps:
Note: Before using for the first time, add the specified volume of anhydrous ethanol to the Washing Buffer RW, mix thoroughly. After adding, please tick the box promptly to mark that ethanol has been added to avoid repeated addition!
1.Transfer 200 μL of body fluids such as serum (which needs to be returned to room temperature; if the volume is insufficient, make up with 0.9% NaCl or PBS) into the aforementioned 1.5 mL centrifuge tube, add 400 μL of Lysis Buffer VLB, and immediately vortex oscillate to mix thoroughly.
If the sample processing volume is small or the expected viral concentration is low, it is recommended to add 4 μL of Poly Carrier stock solution to 400 μL of Lysis Buffer VLB.
2.Place at room temperature (15 - 25℃) for 10 minutes, and oscillate to mix once every 5 minutes.
3.Add 450 μL of anhydrous ethanol, and immediately vortex oscillate to mix thoroughly. If the ambient temperature is higher than 25℃, pre - cool the ethanol on ice before adding.
4.Add the above mixture to an adsorption column SpinRA (place the adsorption column into 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, the solution can be added to the same adsorption column SpinRA 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 Washing Buffer RW (please first check whether anhydrous ethanol has been added!), centrifuge at 12,000 rpm for 30 seconds, discard the waste liquid, and repeat this step by adding another 500 μL of Washing Buffer RW.
7.Put the adsorption column SpinRA back into the empty collection tube, centrifuge at 13,000 rpm for 2 minutes to remove the washing buffer as much as possible, so as to prevent the residual ethanol in the washing buffer from inhibiting subsequent reactions.
8.Take out the adsorption column SpinRA, place it into an RNase - free centrifuge tube, add 30 - 50 μL of RNase - free H₂O (preheating in a 65 - 70℃ water bath in advance will yield better results) to the middle part of the adsorption membrane, place at room temperature for 1 minute, and centrifuge at 12,000 rpm for 1 minute. If a larger amount of DNA/RNA is desired, the obtained solution can be re - added to the spin adsorption column and centrifuged at 12,000 rpm for 1 minute.
The larger the elution volume, the higher the elution efficiency. If a higher concentration of DNA/RNA is required, the elution volume can be appropriately reduced, but the minimum volume should not be less than 20 μL. Too small a volume will reduce the elution efficiency and decrease the DNA/RNA yield.
Viral DNA can be stored at 2 - 8℃. For long - term storage, it can be placed at - 20℃. It is recommended to use viral RNA immediately; otherwise, store it at - 70℃ for short - term use promptly.
