Virus Nucleic Acid Purification Reagent Kit

1. Components of the reagent kit

2. Instructions for Use:

1. The Virus Genome Purification Reagent Kit is intended for molecular biology research and should not be used for the prevention and treatment of diseases.
2. Prior to chemical handling, it is recommended to wear protective clothing, disposable gloves, and safety goggles.
3. Before using Rinse Buffer 1, add (96%~100%) ethanol.
4. The color change to yellow in Reagent Buffer C does not affect normal use.
5. Maintain the centrifugation environment at room temperature (15℃~25℃).

3. Reagent Kit Introduction:

The Virus Genome Purification Reagent Kit provides a rapid and effective solution for virus genome purification, widely applied to various viral nucleic acid purifications, such as Hepatitis B virus, and more.

The purification process can be completed within 1 hour, and the purified nucleic acids can be directly used for PCR, Southern blotting, etc. The entire purification process does not require toxic reagents such as phenol and chloroform, making the nucleic acid purification kit suitable for various other samples.

The purified RNA, washed with a low-salt solution or water, is suitable for downstream experiments. The A260/A280 ratio of the purified RNA is around 1.9, indicating high purity and a well-defined peak at 260 nm.

The high-efficiency membrane effectively removes inhibitors or other interfering substances, making it widely applicable to downstream experiments such as PCR, RAPD, AFLP, RFLP, SNP, and more.

4. Self-provided Equipment and Consumables:

High-speed centrifuge, EP tubes, vortex oscillator, absolute ethanol, hot water bath or metal bath

5.  Extraction Steps:

Preparation before starting:

A. Heat Elution Buffer to 65℃ before use (recommended);

B. Add ethanol toWashBuffer 1 before use.

1. Sample Preparation: Transfer 200μl of serum or other body fluids to a centrifuge tube. If insufficient, top up with PBS.
2. Add 400μl of Reagent Buffer C, vortex thoroughly, let it stand at room temperature for 10 minutes, vortex 4-5 times during this period.
3. Add 450μl of pre-cooled absolute ethanol, vortex immediately.
4. Transfer the obtained liquid to the RNA extraction purification column (set) (approximately 650~700μl each time), let it stand at room temperature for 2 minutes, centrifuge at more than 8,000 rpm for 1 minute, discard the collected waste, and re-insert the collection tube into the purification column for the next step
5. Place the RNA extraction purification column (set) in a new collection tube, add 300μl of Wash Buffer 1, centrifuge at more than 8,000 rpm for 1 minute, discard the waste, and re-insert the RNA extraction purification column (set) into the tube for the next step.

(Note: Confirm that absolute ethanol has been added to Wash Buffer 1.)

6. Add 700μl of Rinse Buffer 1 to the RNA extraction purification column (set), centrifuge at 14,000 rpm (20,000×g) for 2 minutes, extend the centrifugation time appropriately to ensure the membrane is more dry.

(Note: The presence of ethanol has a serious impact on subsequent experiments, so drying the membrane is crucial. After centrifugation, ensure no ethanol is present before elution, then discard the waste and collection tube.)

7. Place the RNA extraction purification column (set) into a new centrifuge tube, drip 100μl of elution buffer onto the membrane, incubate at room temperature for 5 minutes (15℃~25℃), centrifuge at more than 8,000 rpm for 1 minute.

(Note: Eluting with 50μl of elution buffer can increase nucleic acid concentration but reduces total nucleic acid yield.)

8. Repeat the previous step.

(Note: A new centrifuge tube can be used to collect the nucleic acid eluted in the second round, or continue using the original collection tube to collect nucleic acid.)