Virus DNA/RNA Extraction Kit

Virus DNA RNA Extraction Kits

A box of nucleic acid extraction reagent includes Lysis Buffer, Protease K(PK), Wash Buffer I, Wash Buffer II, RElution Buffer, Spin Columns, which is suitable for samples of whole blood, serum, plasma, disease material, feces and body fluids.

1. Simple and fast: the whole DNA RNA extraction process is completed within 30min;
2. High sensitivity: virus DNA  can reach 10 IU / mL, virus RNA can reach 100 IU / mL;
3. Wide application: It can easily extract sputum, saliva, alveolar lavage fluid, (nasopharyngeal) swabs, cultured cells and other body fluid samples (such as whole blood, plasma, serum, ascites, cerebrospinal fluid, oral fluid, etc.), applicable For downstream NGS, qPCR detection;
4. Stable and reliable: The virus DNA extraction kits or RNA extraction kits are no toxic reagents such as phenol and chloroform in the product, and the samples can be stored at room temperature;
5. High purity: The purified viral nucleic acid does not contain contaminants such as PCR / RT-PCR inhibitors.

Principle of Applying DNA RNA Extraction Kits

Lysis is performed in the presence of Protease K and Lysis Buffer, which together ensure the inactivation of Virus DNA and RNA to prepare the virus samples for testing. Binding conditions are adjusted by adding ethanol to allow the optimal binding of the viral RNA and DNA to the membrane. Lysates are then transferred onto a spin column and viral nucleic acids are adsorbed onto the silica-gel membrane as the lysate is drawn through by centrifugation. Nucleic acids remain bound to the membrane, while contaminants are efficiently washed away during 3 wash steps. In a single step, highly pure viral RNA and DNA are eluted in Elution Buffer at room temperature.

Influenza Virus Samples DNA/RNA Extraction Purification Test Procedures

1. Sample pretreatment

Animal/Plant Tissue: Grind sample fully with normal saline or PBS, take the supernatant after centrifugation.

Serum, Plasma, Ascites and other liquid samples: Extraction directly.

Fecal: Add an appropriate amount of normal saline or PBS into the samples, shake them thoroughly, centrifuge them at 12000g for five min, and take the supernatant for extraction.

2. Virus Sample extraction operation

• Pipet 10μL PK into a 1.5mL microcentrifuge tube (not provided).
• Add 200μL sample (If the sample volume ≤200μL, replenish PBS or normal saline to a volume of 200μL) into the microcentrifuge tube.
• Add 200μL Lysis Buffer. Vortex mixing 30seconds.
• Incubate at 56°C for 15min in a heating block. Briefly centrifuge the 1.5mL tube to remove drops from the inside of the tube lid.
• Add 250μL of ethanol (96–100%) to the sample, close the cap and mix thoroughly by pulse-vortexing for 15s. Briefly centrifuge the 1.5mL tube to remove drops from the inside of the tube lid.
• Transfer the mixture into a Spin Column, centrifuge at 10,000g for one minute and discard the flow-through.
• Add 500μL Wash Buffer I into the Spin Column, centrifuge at 10,000g for one minute and discard the flow-through.
• Add 500μL Wash Buffer II into the Spin Column, centrifuge at 10,000g for oneminute and discard the flow-through.
• Add 500μL Wash Buffer II into the Spin Column, centrifuge at 10,000g for oneminute and discard the flow-through.
• Place the spin column in a clean 1.5mL collection tube. Centrifuge at 10,000 g for two min to dry the membrane completely.
• Place the spin column in a clean 1.5mL collection tube. Add 50-100μL Elution Buffer (or RNase-free water pH>7.0) to the center of the membrane; Incubate at the room temperature for twominutes.
• Centrifuge at 12,000g for oneminute. Remove the Spin Basket and discard. Then the buffer in the microcentrifuge tube contains the DNA/ RNA.

Notice

1. Lysis Buffer may be precipitated at low temperature, please heated at 56 ℃ for a few minutes to restore the clarification.
2. Wash Buffer I and Wash Buffer II add the absolute ethanol as the volume marked on the bottle label and mix well

DNA RNA Extraction Kits in Real Clinical Trials for Flu Virus Detection

The virus DNA extraction kits or RNA extraction kits are widely used in labrotary or hospital for medical test use. This virus DNA RNA extraction or purification reagents kits can be used in extracting RNA of different influenza or flu virus for lab tests. The virus samples can be  body fluid from nasal oral or throat, fecal or blood with proper storage solution for its stability for later medical tests. The following three virus detection test procedure and detection data are extracted from our Clinical Trials on positive virus samples using our DNA RNA Extraction Kits for virus RNA extraction.

1. Viral samples from nasal or oral pharyngeal swabs positive for influenza virus at different concentrations were used as samples, and the samples were stored in the virus transpport medium preservative fluid to test the storage stability at 4 ° C and 37 ° C for 7 days. After extracting RNA using viral DNA / RNA extraction or purification Kit II (Cat # BSC71), real time RT-PCR method was used to detect the average Ct comparison results as follows-A human infuenza A serum samples test data for 7 days:

2. Taking fecal coronavirus positive stool as a sample, use the sample preservative solution at a ratio of 1:10 (1g of stool is stored in 10ml of storage solution), compare the storage stability at 4 ° C and 37 ° C for 7 days, and use virus DNA / RNA eatraction purification kit  II (BSC71) for extraction of Fcov-RNA, Real time RT-PCR method is used to detect, and the results of Ct average comparison are as follows:

3. Positive swine whole blood of different concentrations of African swine fever virus (ASFV) was used as a sample, and the sample storage solution was used to store the sample, the storage stability at 37 ° C for 30 days was tested, and the virus RNA extraction purification reagent kits was used after the ASFV-DNA extraction in Box II (Cat # BSC71) for rna extraction for test, real time RT-PCR method was then used to detect the results of Ct average comparison as follows:

Ps. All procedures of applying DNA RNA Extration kits for virua detection need to conducted by medical professionals. Continue to explore more Medical equipments here at Ownhealthylife.