Our Swab/Saliva Viral DNA/RNA Extraction Kit (Spin Column) is suitable for rapid extraction of high purity viral nucleic acid from plasma, serum, nasopharyngeal swab, sputum, bronchoalveolar lavage fluid, ascites, culture cell supernatant and urine. This kit is based on the silica gel column purification method. The sample is homogenized in the lysis buffer and the nucleic acid is released into the buffer. The lysis buffer contains a high concentration of guanidine. In this condition, the membrane absorbs nucleic acid by hydrogen bonding and electrostatic physical and chemical action, while proteins and other impurities are not absorbed. The lysate is transferred to the adsorption column for filtration, and the nucleic acid filter membrane is washed to remove residual proteins and other impurities, and finally eluted by the low-salt buffer solution. The obtained nucleic acids can be directly used in downstream related experiments such as reverse transcription, PCR, RT-PCR, fluorescence quantitative PCR, second-generation sequencing and Northern hybridization.
Main Components
Our kit consists of the following components:
Reagent
Amount
Component Description
Lysis Buffer
50 ml
Provides environment for lysing and binding to the column
Wash Buffer
24 ml
Removes residual proteins and other impurities
Eluent Buffer
6 ml
Nuclease-free solution
Spin Column
50 pieces x 2
Adsorb viral nucleic acid and collect the filtrate
Storage Conditions
Store at room temperature (15-25°C), and transport at RT.
Notes
1. Prepare your own RNase-free pipette tips, 1.5 ml RNase-free microcentrifuge tubes, centrifuge, etc.
2. If the virus has a strong ability to infect, a variety of defense measures must be done before the operation.
3. Avoid repeated freezing and thawing of samples, otherwise the extracted viral RNA will be degraded and the extracted amount will decrease.
4. All operating procedures, if not specified, are carried out at room temperature (15-25°C).
5. When using this kit, please a wear lab coat, disposable latex gloves, disposable masks and use RNase-free consumables to avoid RNase pollution.
6. Please check whether there is crystal precipitation in the Lysis Buffer. If there is crystal precipitation, place it at room temperature or 37°C until the crystal is dissolved. Mix well before use.
Before use
Add 96 ml of 100% ethanol to the Wash Buffer and store at RT.
Protocol
1. Add 500 μl of Lysis Buffer to a 1.5 ml RNase-free microcentrifuge tube (self-provided). If there are many samples, the Lysis Buffer can be pre-packed.
2. Add 200 μl of sample and mix thoroughly by vortexing (if the sample is less than 200 μl, fill it with normal saline to 200 μl).
3. Transfer the above mixture to the Spin Column (with Collection Tube). Centrifuged at 12,000×g for 1 min
4. Discard the filtrate and put the Spin Column back into the 2 ml Collection Tube. Add 600 μl of Wash Buffer and centrifuge at 12,000×g for 30 sec, then discard the filtrate.
Note: make sure the correct amount of anhydrous ethanol has been added to the Wash Buffer.
5. Repeat step 4 once.
6. Centrifuge for 2 min at 12,000×g to dry the column membrane. Discard the filtrate and collection tube.
7. Transfer the Spin Column to a new 1.5 ml Collection Tube (provided by the kit), add 50 μl Eluent Buffer to the center of the membrane of the Spin Column, and place it at room temperature for 1 min. Centrifuge at 12,000×g for 1 min.
8. Discard the Spin Column, the obtained DNA/RNA can be directly used for subsequent detection, or be stored at -30 to -15°C for short-term storage or at -70°C for long-term storage.
