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Total RNA Extraction Kit (Trizol)
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AN51L758100mL298
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Product Description
The main component of total RNA extraction reagent (Trizol; hereafter referred to as “Trizol”) is guanidinium isothiocyanate, which disrupts cells and releases cellular RNA while preserving RNA integrity. Consequently, it is widely used for extracting RNA from both animal and plant cells.
Order Information
Product Name
Item number
Specifications
Total RNA extraction reagent (Trizol)
AN51L758
100mL
Transportation and Storage
Store at room temperature. Protect from light and keep at 4°C; shelf life is 12 months.
Instructions for Use
1. Prepare the sample
l Tissue samples: Use liquid nitrogen–assisted grinding. Place tissue pieces directly into a mortar, add a small amount of liquid nitrogen, and grind rapidly. Once the tissue softens, add a further small amount of liquid nitrogen and continue grinding. Repeat this process three times, then add Trizol at a ratio of 50–100 mg/mL per milligram of tissue. Homogenize. [Note]: The volume of tissue should not exceed 10% of the Trizol volume; otherwise, homogenization will be inefficient. When using an electric homogenizer, homogenization typically takes about 1–2 minutes.
l Cells: For adherent cells, no enzymatic digestion is required; they can be directly subjected to Trizol‑based lysis. Add Trizol at a ratio of 10 cm² per mL. Suspension cells can be collected directly, followed by enzymatic digestion and lysis; each mL of Trizol can lyse approximately 5 × 10⁶ animal, plant, or yeast cells, or 1 × 10⁷ bacterial cells.
2. After adding Trizol to cells or tissues, allow them to stand at room temperature for 5 minutes to ensure complete lysis. [Note]: At this point, the samples can be stored at −70°C for long-term preservation.
3. Centrifuge at 12,000g for 5 min, and discard the pellet.
4. Add chloroform at a ratio of 200 μL chloroform per mL Trizol, shake vigorously to mix, and let stand at room temperature for 15 minutes. [Note]: Do not use a vortex mixer to avoid shearing genomic DNA.
5. Centrifuge at 4°C at 12,000g for 15 min.
6. Carefully transfer the upper aqueous phase into another centrifuge tube. [Note]: Do not under any circumstances pipette the interphase. If both DNA and protein are being extracted, retain the lower phenol phase and store it at 4°C; if only RNA is being isolated, discard the lower phenol phase.
7. Add 0.5 mL of isopropanol to each mL of TRIzol, mix thoroughly, and let stand at room temperature for 5–10 minutes.
8. Centrifuge at 4°C at 12,000g for 10 min; discard the supernatant, leaving the RNA precipitated at the bottom of the tube.
9. Add 1 mL of 75% ethanol to the trizol, gently vortex the centrifuge tube, and resuspend the precipitate.
10. Centrifuge at 4°C at 8,000g for 5 min, and carefully discard the supernatant.
11. Allow to air-dry at room temperature or dry under vacuum for 5–10 minutes. [Note]: Do not over‑dry the RNA sample, as it will be difficult to dissolve otherwise.
12. Available in 50 μL H 2 Dissolve the RNA sample in O, TE buffer or 0.5% SDS at 55–60°C for 5–10 minutes. [Note]: H 2 O, TE buffer, or 0.5% SDS must all be treated with DEPC and autoclaved.
Precautions
1. This product is intended solely for scientific research and must not be used for clinical diagnosis, treatment, or any other such purposes.
2. Operators shall undergo specialized training and strictly adhere to operating procedures.
3. Handling and operations shall be conducted in areas equipped with local exhaust ventilation or general ventilation systems.
4. Avoid contact with eyes and skin, and avoid inhaling vapors.
5. Keep away from sources of ignition and heat; smoking is strictly prohibited in the workplace.
6. The entire RNA extraction process requires RNase-free laboratoryware.
Related Product Recommendations
RNA Extract auxiliary reagent (chloroform substitute) ( Item number :AN51L675)
Total RNA Column-Based Extraction Kit ( Item number :AN51L517)
This product is intended solely for scientific research and must not be used for clinical diagnosis, treatment, or any other purposes.
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