7. Virus DNA/RNA Extraction Kit, 50 T $199,

Description

Product Introduction

This kit is designed for the rapid and convenient extraction of various virus DNA/RNA from serum, plasma, urine, cell culture supernatant, virus stock, and infected virus tissues. By utilizing the reversible binding property of nucleic acids to the nucleic acid binding column, combined with a specialized buffer system, this product allows specific binding of nucleic acids to the membrane matrix while effectively washing away proteins, cell debris, and other contaminants. Finally, the DNA/RNA is eluted from the binding column using Nuclease-free Water. The obtained high-quality DNA/RNA can be directly used for downstream molecular biology experiments such as PCR, cDNA synthesis, and RT-qPCR.

Storage and Transportation

Proteinase K and Carrier RNA are transported in ice bags (wet ice) and should be stored at -20°C. The remaining reagents are transported and stored at room temperature. The shelf life is 12 months.

Composition

Preparation Before Use

1. If Buffer VLB and Buffer VWA show precipitation, heat them at 37°C to dissolve and allow them to return to room temperature before use.
2. Before the first use, add 18 mL of nuclease-free ethanol to Buffer VWA and 60 mL of nuclease-free ethanol to Buffer VWB.
3. Carrier RNA can be aliquoted and stored at -20°C before the first use. Avoid repeated freeze-thaw cycles.
4. Pre-cool the tissue grinder in advance if using one.

Experimental Procedure

1. Handling of Virus Samples:

a) Lysis of serum, plasma, urine, cell culture supernatant, and virus stock: Take 10-200 μL of serum, plasma, urine, cell culture supernatant, or virus stock. If the initial volume is less than 200 μL, add PBS or nuclease-free water to reach a volume of 200 μL.

b) Lysis of infected virus tissue: Take 10-20 mg of fresh or ultra-low temperature frozen infected virus tissue. Place it in a 1.5 mL nuclease-free centrifuge tube or a dedicated grinding tube (recommended: HT-200-M) containing 2-3 3 mm grinding beads (recommended: G0203). Immediately place the tube with the tissue in liquid nitrogen, then use a tissue grinder (recommended: KZ-5F-3D) to thoroughly grind the tissue to a homogenized state under low-temperature conditions (incomplete homogenization will affect DNA/RNA yield and quality). After grinding, add 200 μL of PBS or nuclease-free water.

2. Add 200 μL of Buffer VLB, 20 μL of Proteinase K, and 3 μL of Carrier RNA. Mix thoroughly and incubate at 56°C for 10 min.
3. Add 200 μL of nuclease-free ethanol and mix by inverting the tube.
4. Transfer the solution from the previous step to a spin column. Centrifuge at 12,000 rpm for 1 min and discard the filtrate in the collection tube.
5. Add 500 μL of Buffer VWA to the spin column. Centrifuge at 12,000 rpm for 1 min and discard the filtrate in the collection tube.
6. Add 700 μL of Buffer VWB to the spin column (adding Buffer VWB along the wall of the spin column helps wash away residual salts on the column walls). Centrifuge at 12,000 rpm for 30 s and discard the waste liquid.
7. Repeat step 6.
8. Place the spin column on a collection tube and centrifuge at 12,000 rpm for 2 min.
9. Transfer the spin column to a new nuclease-free 1.5 mL centrifuge tube, open the lid, and let it stand at room temperature for 3-5 min to allow the residual ethanol on the spin column to evaporate completely.
10. Add 30-50 μL of nuclease-free water to the spin column. Let it stand at room temperature for 5 min. Centrifuge at 12,000 rpm at room temperature for 2 min to elute the DNA/RNA. If a higher concentration of DNA/RNA is desired, the first elution solution can be added back to the spin column, let it stand at room temperature for 5 min, centrifuge at 12,000 rpm at room temperature for 2 min, and collect the DNA/RNA again.

Notes

1. Please read the product manual carefully before operation.
2. Due to the addition of Carrier RNA during the extraction