The Yeasen Exosome RNA Isolation Kit employs a phenol/guanidine-based method to extract total RNA (including miRNA) from isolated exosomes. It efficiently releases RNA from exosomes obtained via ultracentrifugation, chemical precipitation, immunocapture, or size exclusion chromatography. Using a spin column with specific nucleic acid adsorption properties, the kit allows for convenient and rapid purification and elution of exosomal RNA. The purified exosomal RNA is directly suitable for downstream applications such as RT-qPCR, Northern blot, microarray expression profiling, and NGS sequencing.

Product Information

Components

Storage

Ship with ice packs. Store at 2-8°C (components should be stored according to the conditions specified above). Valid for 1 year.

Instructions

1. Exosome RNA Release

Add 200 μL of the isolated exosome solution to 200 μL of Lysis Buffer A. Vortex vigorously for 30 sec and incubate at room temperature for 5 min to ensure complete lysis of exosomes.

2. Exosome RNA Separation

Add 200 μL of Lysis Buffer B, cap the tube, and shake vigorously for 15 sec. Incubate at room temperature for 5 min. Then add 200 μL of Lysis Buffer C, cap the tube, and shake vigorously for 15 sec. Incubate at room temperature for 5 min. Centrifuge at 4°C, 12,000×g for 15 min. The sample will separate into three phases: a lower yellow organic phase, an interphase, and an upper colorless aqueous phase. The RNA is primarily located in the upper aqueous phase. Carefully transfer the aqueous phase to a spin column (avoiding the interphase) for the next step.

3. Exosome RNA Washing
3.1 Let the supernatant stand in the column for 5 min. Centrifuge at 4°C, 8,000×g for 2 min. Discard the liquid in the collection tube.
3.2 Add 500 μL of Washing Buffer W to the column. Let it stand for 5 min, then centrifuge at 4°C, 8,000×g for 2 min. Discard the liquid in the collection tube.
3.3 Empty column: Centrifuge at 4°C, 12,000×g for 2 min to remove residual liquid. Discard the collection tube and transfer the spin column to a new 1.5 mL centrifuge tube (provided in the kit). Open the column lid and let it stand at room temperature for several minutes (5-10 min) to completely dry any residual washing buffer from the adsorption matrix.

4. Exosome RNA Elution

Pipette 5 Boiler Buffer E onto the center of the column membrane. Incubate at room temperature for 2-5 min. Centrifuge at 4°C, 12,000×g for 2 min to collect the solution in the 1.5 mL centrifuge tube.

Notes

1. Before use, check for salt precipitation in the reagents. If precipitation occurs, incubate at 37°C for 10 min, mix well until no precipitate remains and the solution is clear.

2. To prevent RNase contamination, use RNase-free plasticware and pipette tips to avoid cross-contamination. Prepare solutions using RNase-free water.

3. For your safety and health, please wear a lab coat and disposable gloves during operation.

4. This product is for scientific research use only!

Documents:

Safety Data Sheet

41228_MSDS_HB260525

Manuals:

41228_Manual_HB20260518

FAQs

Q1: At what temperature should centrifugation be performed when extracting exosomal RNA?

A: All centrifugation steps should be performed at 4°C.

Q2: Why is the A260/280 ratio of the extracted exosomal RNA lower than 2.0?

A: Most exosomal RNA consists of small RNAs and has a low concentration. The A260/280 ratio tends to decrease with lower RNA concentrations. A normal A260/280 ratio for exosomal RNA ranges from 1.0 to 1.6. A low ratio will not affect downstream applications.

Q3: Why is the performance in downstream applications (e.g., RT-PCR) not optimal?

A: Ethanol and/or excess salt ions in the eluted exosomal RNA may inhibit PCR reactions. Therefore, after the final washing step, ensure the column is placed at room temperature for several minutes to completely dry any residual washing buffer.

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