Products

Product Introduction:
This kit adopts an improved guanidinium isothiocyanate/phenol one-step method to lyse cells and inactivate RNases. Total RNA then selectively binds to the silica matrix membrane inside the spin column under high chaotropic salt conditions. Through a series of rapid washing-centrifugation steps, deproteinization buffer and washing buffer remove impurities such as cellular metabolites and proteins. Finally, pure RNA is eluted from the silica matrix membrane using low-salt RNase-free water.

Product Features:
1.The silica matrix membranes in the spin columns are all high-quality specially modified membranes, ensuring minimal variation in adsorption capacity between columns.
2.Combines the advantages of guanidinium isothiocyanate/phenol one-step reagents (excellent stability and high purity) and spin columns (convenience and speed). No isopropanol precipitation or ethanol washing steps are required; RNA can be directly eluted from the spin column, avoiding the problem of difficulty in dissolving due to over-drying.
3.The unique formulation of Buffer RLS lysis buffer allows direct lysis of whole blood without prior red blood cell removal.
4.Multiple deproteinization washing steps ensure higher purity of the extracted RNA.
5.Effectively reduces the content of 5S RNA in total RNA, improving purity.

Application Scope:
Suitable for rapid extraction of high-purity total RNA from whole blood (liquid samples).

Operation Steps:
Note: Before first use, add the specified volume of anhydrous ethanol to the Washing Buffer WB bottle, mix thoroughly. Immediately mark the corresponding checkbox after adding ethanol to avoid repeated addition!

1.Add 0.75 mL Lysis Buffer RLS to 0.25 mL liquid sample (serum, plasma, cerebrospinal fluid, etc.). Pipette the liquid sample several times to assist in lysing cells within the sample. Add at least 0.75 mL Lysis Buffer RLS per 5~10×10⁶ cells. The final volume ratio of Lysis Buffer RLS to liquid sample must always be 3:1.

2.Vigorously vortex the sample to mix well, and incubate at 15-30°C for 5 minutes to fully dissociate ribosomes.

3.Add 0.2 mL chloroform to 0.75 mL Lysis Buffer RLS, vortex vigorously for 15 seconds, and incubate at room temperature for 2 minutes.

4.Centrifuge at 12,000 rpm for 10 minutes at 4°C. The sample will separate into three layers: the lower organic phase, the interphase, and the upper colorless aqueous phase. RNA is present in the aqueous phase. The volume of the aqueous phase is approximately 70% of the added RLS volume. Transfer the aqueous phase to a new tube and proceed to the next step.

5.Add an equal volume of 70% ethanol (please check if anhydrous ethanol has been added first!), invert to mix (precipitation may form at this step). Transfer the resulting solution and any precipitate to a SpinRA RNA Spin Column (place the column in a collection tube).

6.Centrifuge at 12,000 rpm for 45 seconds, discard the waste liquid, and place the spin column back into the collection tube.

7.Add 500 μL of Deproteinization Buffer RE, centrifuge at 12,000 rpm for 45 seconds, and discard the waste liquid.

8.Add 500 μL of Washing Buffer RW (please check if anhydrous ethanol has been added first!), centrifuge at 12,000 rpm for 45 seconds, and discard the waste liquid.

9.Add another 500 μL of Washing Buffer RW, centrifuge at 12,000 rpm for 45 seconds, and discard the waste liquid.

10.Place the SpinRA RNA Spin Column back into an empty collection tube, centrifuge at 13,000 rpm for 2 minutes to thoroughly remove residual washing buffer, as ethanol residues in the washing buffer may inhibit downstream reactions.

11.Remove the SpinRA RNA Spin Column and place it into an RNase-free centrifuge tube. Add 30-50 μL of RNase-free water to the center of the adsorption membrane (preheating the water in a 65-70℃ water bath in advance yields better results) based on the expected RNA yield. Incubate at room temperature for 2 minutes, then centrifuge at 12,000 rpm for 1 minute. For higher RNA yield, load the obtained solution back into the 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.

Note: Larger elution volume results in higher elution efficiency. If higher RNA concentration is required, the elution volume can be appropriately reduced, but the minimum volume should be no less than 30 μL. Excessively small volume reduces RNA elution efficiency and yield.