RNA clean-up and concentration kit, using Tini spin column, 50 preps

Feature:

- Easy and rapid with 10 min. procedure using RNA Tini spin column.

- Nearly 100% RNA recovery

- Cleanup RNA from mini prep and after enzymatic reactions

- Concentrate RNA to as low as 5μl volume

- Remove Salt, primers, enzymes and other impurities

Kit Contents:

Components RNA Cleanup and Concentration (Cat#702)

RLT Solution*: 14ml

RW Solution: 12ml

RPE Solution**: 5ml (add 20ml 100% ethanol to the bottle before use)

RNase-free water: 1.5ml

Tini spin column with collection tube: 50 sets

*RLT Solution contains chaotropic salt. Harmful by inhalation, in contact with skin and if swallowed (Risk and Safety

phrases: R20/21/22-32, S13-26-36-46)

**Add 100% ethanol before use: add 20ml 100% ethanol to 5ml RPE Solution.

Storage: Store all Buffers at 4oC for up to 2 years.

Note: In general, DNase digestion is not required since the RNeasy silica-membrane technology efficiently removes most of the DNA without

DNase treatment. However, further DNA removal may be necessary for certain RNA applications that are sensitive to very small amounts of

DNA (e.g., TaqMan RT-PCR analysis with a low-abundant target). DNA can be removed by a DNase digestion following RNA isolation.

Protocol:

1. Adjust sample to a volume of 50μl with RNase-free H₂O, add 175μl of RLT Solution, and mix well. Add

130μl of 100% ethanol, mix gently. A precipitate may form by adding ethanol, but this will not affect the procedure.

Note: for clean-up micro RNA (17nt-200nt), add more ethanol to 60% final concentration, eg: 50μl RNA

sample+175μl RLT solution+350μl 100% ethanol. Precipitate may form after adding ethanol, but this will

not affect the procedure. Do not load more than 400μl of sample on Tini spin column at one time.

Discard the flow-through and load more sample mixture if needed.

2. Place a Tini Spin Column in 1.5ml Collection Tube and transfer the mixture (Step 1) to the column and

spin at full speed ~10,000 rpm (12,000 x g) for 1 minute, discard flow-through.

3. Add 200μl of RW Solution to the column and spin at ~10,000 rpm (12,000 x g) for 1 minute to remove

small RNA (<200 nt). (Skip this step if you want to recover micro RNA)

4. Add 200μl of RPE Solution to the column and spin at ~10,000 rpm (12,000 x g) for 1 minute, discard the

flow-through and spin once more to completely remove the residue of RPE Solution.

Important: residual ethanol from RPE Solution may affect downstream applications.

5. Add 5-20μl of RNase-free H2O onto the center of the column membrane and centrifuge at 10,000 rpm for

1 minute. Keep RNA sample at -70℃.

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