Products

Product Description:
This kit uses spin columns packed with specially imported silica matrix membranes. These membranes ensure minimal variation in RNA binding capacity between columns and excellent reproducibility.RNA binds efficiently and specifically to the silica matrix membrane under high-salt conditions. Meanwhile, most proteins, inorganic ions, and many organic impurities are removed. RNA is eluted under low-salt conditions.The kit can process RNA samples up to 50 μg. It is designed for purifying and recovering RNA from enzyme reaction mixtures, such as DNase-treated, protease-treated, and RNA-labeled samples. It is also suitable for purifying RNA extracted by other methods.The purified total RNA is free of protein contamination. It is suitable for downstream applications including Northern blot, Dot blot, mRNA isolation, cDNA synthesis, primer extension, and differential display.

Operating Procedures:
Note: Before first use, add the specified volume of anhydrous ethanol to Washing Buffer RW and mix thoroughly. Mark the box immediately after addition to avoid repeated ethanol addition!
All the following steps can be performed at room temperature. Operate quickly to minimize RNA degradation.

1.Place the RNA sample on ice. Add RNase-free water to adjust the total volume to 100 μL. Add 350 μL of Binding Buffer RC and mix well.

2.Add 250 μL of anhydrous ethanol and mix well. No centrifugation is required.

3.Transfer the resulting solution, along with any potential precipitates, into the Spin Column RA (placed inside the collection tube). Centrifuge at 12,000 rpm for 45 seconds at 4℃. Discard the waste liquid in the collection tube and place the spin column back into the tube.
To remove trace DNA residues, perform on-column DNase digestion after this step. See the Appendix for detailed procedures.

4.Add 0.5 mL of Washing Buffer RW (check ethanol addition beforehand). Centrifuge at 12,000 rpm for 45 seconds at 4℃. Discard the waste liquid.

5.Add another 0.5 mL of Washing Buffer RW. Centrifuge at 12,000 rpm for 45 seconds at 4℃. Discard the waste liquid.

6.Centrifuge at 13,000 rpm for 2 minutes at 4℃ to completely remove residual washing buffer. This step prevents residual ethanol in the buffer from inhibiting downstream reactions.

7.Remove the Spin Column RA and place it into a new RNase-free centrifuge tube. Add 50–80 μL of RNase-free water (preheated in a 65–70℃ water bath for better results) directly onto the center of the silica membrane. Incubate at room temperature for 2 minutes, then centrifuge at 12,000 rpm for 1 minute.For higher RNA yield: Reapply the eluate to the same spin column and centrifuge for 1 minute, or add an additional 30 μL of RNase-free water, centrifuge for 1 minute, and combine the two eluates.
Larger elution volumes improve elution efficiency. For higher RNA concentration, reduce the elution volume appropriately, but do not use less than 30 μL, as volumes below this will decrease elution efficiency and RNA yield.