Products

Product Introduction:
The silica matrix membrane in the spin column of this kit is a special imported adsorption membrane, with minimal differences in adsorption capacity between columns and good reproducibility. Under high-salt conditions, RNA binds efficiently and specifically to the silica adsorption membrane, while maximizing the removal of proteins, inorganic salt ions, and many organic impurities. Under low-salt conditions, RNA is eluted. The kit can process up to 50 μg of RNA samples. It is used for the purification and recovery of RNA from enzyme reaction solutions (such as DNase treatment, Proteinase treatment, RNA labeling, etc.), and can also be used for the purification of RNA obtained by other extraction methods. The purified total RNA is free of protein contamination and can be used for Northern blot, Dot blot, mRNA extraction, cDNA synthesis, primer extension, differential display, and other experiments.

Experimental Procedures:
Notes
Before first use, add the indicated amount of absolute ethanol to the Wash Buffer RW bottle, mix thoroughly, and mark with a check immediately after adding to avoid repeated addition!
All the following steps can be performed at room temperature, but operations should be carried out quickly to reduce the chance of RNA degradation.

1.On ice, add RNase-free water to the RNA sample to make up to 100 μL, then add 350 μL of Binding Buffer RC and mix well.

2.Add 250 μL of absolute ethanol and mix well; no centrifugation is required.

3.Transfer the solution from the previous step (including any possible precipitation) to a SpinRA column (the spin column is sleeved in a collection tube), centrifuge at 12,000 rpm at 4°C for 45 seconds. Discard the waste liquid in the collection tube and reattach the spin column to the collection tube.
To remove trace DNA residue, on-column DNase digestion can be performed after this step; see the appendix for details.

4.Add 0.5 mL of Wash Buffer RW (please check if ethanol has been added first), centrifuge at 12,000 rpm at 4°C for 45 seconds, and discard the waste liquid.

5.Add 0.5 mL of Wash Buffer RW, centrifuge at 12,000 rpm at 4°C for 45 seconds, and discard the waste liquid.

6.Centrifuge at 13,000 rpm at 4°C for 2 minutes to remove as much wash buffer as possible, avoiding residual ethanol in the wash buffer that may inhibit downstream reactions.

7.Remove the SpinRA column and place it into a new RNase-free centrifuge tube. According to the expected RNA yield, add 50-80 μL of RNase-free water to the center of the adsorption membrane (heating the water in a 65-70°C water bath in advance yields better results). Incubate at room temperature for 2 minutes, then centrifuge at 12,000 rpm for 1 minute. To obtain more RNA, re-add the eluate to the spin column and centrifuge for 1 minute, or add an additional 30 μL of RNase-free water and centrifuge for 1 minute, then combine the two eluates.
The larger the elution volume, the higher the elution efficiency. If a higher RNA concentration is required, the elution volume can be appropriately reduced, but the minimum volume should not be less than 30 μL. Too small a volume will reduce RNA elution efficiency and yield.