Virus Nucleic Acid Purification Reagent Kit

1. Komponen kit reagen

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, menambahkan (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, dan banyak lagi.

The purification process can be completed within 1 jam, and the purified nucleic acids can be directly used for PCR, Blot Selatan, dll.. 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, dan banyak lagi.

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. Persiapan Sampel: Transfer 200μl of serum or other body fluids to a centrifuge tube. If insufficient, top up with PBS.
2. Menambahkan 400μl Reagen Buffer C, vortex thoroughly, let it stand at room temperature for 10 menit, vortex 4-5 kali selama periode ini.
3. Menambahkan 450μl of pre-cooled absolute ethanol, vortex immediately.
4. Transfer the obtained liquid to the Ekstraksi RNA purification column (mengatur) (sekitar 650~700μl setiap kali), let it stand at room temperature for 2 menit, centrifuge lebih dari 8,000 rpm untuk 1 menit, membuang sampah yang dikumpulkan, dan masukkan kembali tabung pengumpul ke dalam kolom pemurnian untuk langkah selanjutnya
5. Place the RNA extraction purification column (mengatur) dalam tabung koleksi baru, menambahkan 300μl of Wash Buffer 1, centrifuge lebih dari 8,000 rpm untuk 1 menit, membuang limbahnya, and re-insert the RNA extraction purification column (mengatur) ke dalam tabung untuk langkah selanjutnya.

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

6. Menambahkan 700μl of Rinse Buffer 1 to the RNA extraction purification column (mengatur), sentrifugasi di 14,000 rpm (20,000×g) untuk 2 menit, extend the centrifugation time appropriately to ensure the membrane is more dry.

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

7. Place the RNA extraction purification column (mengatur) ke dalam tabung centrifuge yang baru, drip 100μl of elution buffer ke membran, incubate at room temperature for 5 menit (15℃~25℃), centrifuge lebih dari 8,000 rpm untuk 1 menit.

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

8. Repeat the previous step.

(Catatan: 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.)