Total RNA Isolation Kit (Tissue)

The Total RNA Isolation Kit (Tissue) provides a fast, simple, and cost-effective method for isolation of total RNA from the tissue sample. Detergents and chaotropic salt are used to lyse cells and inactivate RNase. The specialized high-salt buffering system allows RNA species bases to bind to the glass fiber matrix of the spin column while contaminants pass through the column. Impurities are efficiently washed away, and the pure RNA is eluted with REL Buffer without phenol extraction or alcohol precipitation. RNA purified with The Total RNA Isolation Kit is suitable for a variety of routine applications including RT-PCR, cDNA Synthesis, Northern Blotting, Differential display, Primer Extension and mRNA Selection. The entire procedure can be completed within 25-40 minutes.

• Sample: up to 30 mg of tissue, up to 25 mg of paraffin-embedded tissue
• Format: Spin column
• Operation time: 25-40 minutes
• Elution volume: 50 μl
• Yield: up to 30 μg

Kit Contents

• Delivering high-quality total RNA with the fast procedure
• Ready-to-use RNA for high performance in any downstream application
• Consistent RNA yield from the starting material with a small amount

Sample Preparation

• Fresh or Frozen Tissue

1. Cut off up to 30 mg of fresh or frozen animal tissue and grind the sample under liquid nitrogen to a fine powder using a mortar and pestle. (If using frozen animal tissue, the sample MUST have been flash frozen in liquid nitrogen and immediately stored at -70°C until use, to avoid RNA Degradation).
2. Proceed with the Step2 Lysis.

• Paraffin-embedded tissue

1. Slice small sections (up to 25 mg) from blocks of paraffin-embedded tissue and transfer to a 1.5 ml microcentrifuge tube.
2. Add 1 ml of xylene to the tube.
3. Vortex vigorously and incubate at room temperature for approximately 10 minutes.
4. Vortex occasionally during incubation.
5. Centrifuge at 14-16,000 x g for 3 minutes. Remove the supernatant.
6. Add 1 ml of absolute ethanol to wash the sample pellet and mix by inverting.
7. Centrifuge at 14-16,000 x g for 3 minutes. Remove the supernatant.
8. Add 1 ml of absolute ethanol to wash the sample pellet again and mix by inverting.
9. Centrifuge at 14-16,000 x g for 3 minutes. Remove the supernatant.
10. Open the tube and Incubate at 37°C for 15 minutes to evaporate any ethanol residue.
11. Proceed with the Step 2 Lysis.

Refer to the table below to troubleshoot problems that you may encounter when purifying total RNA.

• Buffers RR and W1 contain irritants. Wear gloves when handling these buffers.
• Add 60 ml of the ethanol(96~100%) to the Buffer W2 before use.
• Check Buffers before use for salt precipitation. Re-dissolve any precipitate by warming up to 37°C.
• During the operation, always wear the latex or vinyl gloves while handling reagents and RNA samples to prevent the RNase contamination.
• Research Use Only. Not intended for any animal or human therapeutic or diagnostic uses.