Kit de reactivos de purificación de ácidos nucleicos de virus

1. Componentes del kit de reactivos

2. Instrucciones de uso:

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, agregar (96%~100%) etanol.
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, y más.

The purification process can be completed within 1 hora, and the purified nucleic acids can be directly used for PCR, transferencia del sur, 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 Nuevo Méjico.

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, y más.

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 (recomendado);

B. Add ethanol toLavarBuffer 1 antes de usar.

1. Preparación de la muestra: Transfer 200μl of serum or other body fluids to a centrifuge tube. If insufficient, top up with PBS.
2. Agregar 400μl of Reagent Buffer C, vórtice a fondo, let it stand at room temperature for 10 minutos, vortex 4-5 times during this period.
3. Agregar 450μl of pre-cooled absolute ethanol, vortex immediately.
4. Transfer the obtained liquid to the extracción de ARN purification column (colocar) (aproximadamente 650~700μl cada vez), let it stand at room temperature for 2 minutos, centrifuge at more than 8,000 rpm para 1 minuto, desechar los residuos recogidos, y vuelva a insertar el tubo de recolección en la columna de purificación para el siguiente paso
5. Place the RNA extraction purification column (colocar) en un nuevo tubo de recolección, agregar 300µl de Lavar Buffer 1, centrifuge at more than 8,000 rpm para 1 minuto, desechar los residuos, and re-insert the RNA extraction purification column (colocar) en el tubo para el siguiente paso.

(Nota: Confirm that absolute ethanol has been added to Lavar Buffer 1.)

6. Agregar 700µl de tampón de enjuague 1 to the RNA extraction purification column (colocar), centrifugar en 14,000 rpm (20,000×g) para 2 minutos, extend the centrifugation time appropriately to ensure the membrane is more dry.

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

7. Place the RNA extraction purification column (colocar) en un nuevo tubo de centrífuga, drip 100µl de tampón de elución sobre la membrana, incubar a temperatura ambiente durante 5 minutos (15℃ ~ 25 ℃), centrifuge at more than 8,000 rpm para 1 minuto.

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

8. Repeat the previous step.

(Nota: 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.)