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exoRNeasy Midi and Maxi Kits

For efficient isolation of exosomal RNA from biofluids

  1. Highly pure total RNA (including the small RNA fraction) from exosomes and other extracellular vesicles (EVs)
  2. RNA isolation exclusively from exosomes, excluding RNA from non-vesicular protein complexes
  3. Convenient on-column procedure provides access to RNA in just one hour
  4. High sensitivity provided by maxi kit that allows large sample input volumes to detect low-abundance RNAs
  5. Isolation from most biofluids, including urine, serum, plasma, CSF and cell culture supernatant

exoRNeasy Kits use membrane affinity spin columns to efficiently capture exosomes and other EVs from most biofluids, including urine, serum, plasma, CSF and cell culture supernatant. The kits then use proven RNeasy MinElute spin columns to isolate RNA from the EVs. The midi column format enables efficient processing of smaller sample volumes (up to 1 ml serum/plasma or 4 ml of urine), while the maxi column format allows the use of large sample volumes, up to 4 ml serum/plasma, 16 ml urine or 32 ml cell culture supernatant, to detect low-abundance RNAs with high confidence. The results are fast, consistent and highly suited to sensitive downstream applications. The RNA isolation part of the protocol using exoRNeasy Kits can be automated on the QIAcube Connect.

Need a quote for your research project or would you like to discuss your project with our specialist team? Just contact us.

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Cat No./ID: 76204
Buffer XBP (250 ml)
$98.00
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250 ml Binding Buffer
Cat No./ID: 76214
Buffer XE (45 ml)
$31.10
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45 ml Elution Buffer for elution of intact extracellular vesicles

Cat No./ID: 77023
exoRNeasy Serum/Plasma Starter Kit (20)
$738.00
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For purification of total exosome-derived RNA from serum/plasma; includes 20 exoEasy columns (10 Maxi, 10 Midi) and 20 RNeasy MinElute spin columns
Cat No./ID: 77144
exoRNeasy Midi Kit (50)
$1,315.00
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For purification of total exosome-derived RNA; includes 50 exoEasy Midi columns and 50 RNeasy MinElute spin columns
Cat No./ID: 77164
exoRNeasy Maxi Kit (50)
$2,691.00
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For purification of total exosome-derived RNA; includes 50 exoEasy Maxi columns and 50 RNeasy MinElute spin columns
Cat No./ID: 77900
Reagent UI (1 ml)
$55.90
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Urine inhibitor removal reagent for use with the exoRNeasy Midi Kit or exoRNeasy Maxi Kit. Contents are sufficient for use with one kit.
exoRNeasy Midi and Maxi Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.

Product Details

0
exoRNeasy captures all mRNA and vesicle-specific miRNAs in plasma.
RNA from pre-filtered plasma was isolated with exoRNeasy and the flow-through of the exoEasy column was used in direct lysis. Shown are raw CT values from RT-qPCR experiments with duplicate replicate isolations and duplicate qPCR reactions.

* Arroyo, J.D. et al. (2011) Argonaute2 complexes carry a population of circulating microRNAs independent of vesicles in human plasma. Proc. Natl. Acad. Sci. USA 108, 5003.
1
exoRNeasy isolates small and large RNAs from extracellular vesicles present in plasma.
Bioanalyzer sizing was performed with vesicle-derived RNA purified by two methods. The plasma was pre-filtered (0.8 µm) to exclude larger particles and subjected to either ultracentrifugation, the current gold standard of vesicle isolation, or the exoRNeasy procedure. Both methods purify RNA of similar size and yield.
2
The exoRNeasy Serum/Plasma Maxi Kit workflow – sample to microvesicles to total RNA, in just one hour.
Starting with filtered plasma, the exoRNeasy Serum/Plasma Maxi Kit provides microvesicle isolation in just 20 minutes. A subsequent 35-minute isolation procedure yields total RNA. Just one hour from sample to microvesicle RNA means you can spend less time getting the RNA, and more time deciphering what it means.
3
Intact vesicles are eluted from the exoEasy membrane with higher purity compared with ultracentrifugation.

Scanning electron microscopy (SEM; 20,000x magnification) was performed on a solubilized pellet from ultracentrifugation of pre-filtered (0.8 µm) plasma compared with a non-lysed eluate of an exoEasy column. Both preparations contain vesicle-shaped structures with an expected size range from 50–200 nm (white arrows; scale bar 200 nm). The preparation from ultracentrifugation also includes many smaller, unidentified structures that do not match the expected shape and size of extracellular vesicles.

Performance

The protocol for exoRNeasy purification of exosomes and other extracellular vesicles is not only faster than ultracentrifugation, but also yields a cleaner preparation. Scanning electron microscopy shows that while both techniques deliver intact vesicles of the expected size, preparation from ultracentrifugation also contains many smaller structures that do not match the expected size or shape for EVs (see figure Intact vesicles are eluted from the exoEasy membrane with higher purity compared with ultracentrifugation).

Moreover, both exoRNeasy and the traditional ultracentrifugation method deliver similar sizes and yields of large and small RNAs, as measured by Bioanalyzer analysis (see figure exoRNeasy isolates small and large RNAs from extracellular vesicles in plasma).

Finally, exoRNeasy captures all cell-free mRNAs in plasma, as well as EV-associated miRNAs, as confirmed by RT-qPCR experiments on the RNAs purified with the exoRNeasy protocol and the flow-through from the exoEasy column (see figure exoRNeasy captures all mRNA and vesicle-specific miRNAs in plasma).

Principle

exoRNeasy Kits improve upon traditional ultracentrifugation microvesicle isolation methods, yielding purified extracellular vesicles in just 20 minutes. Using technology developed with Exosome Diagnostics, Inc., the kits use a spin column format and specialized buffers to capture exosomes from prefiltered biofluids; up to 32 ml using the maxi kit, and 4 ml using the midi kit. Total RNA is then extracted using QIAGEN miRNeasy technology. The entire procedure, from sample to RNA, takes only one hour. 

The exoRNeasy Serum/Plasma Starter Kit provides both midi and maxi exoEasy columns, enabling analysis at varied volumes of prefiltered biofluids.

Procedure

The total procedure for isolating RNA from extracellular vesicles comprises two phases: exosome purification and RNA isolation (see figure The exoRNeasy Serum/Plasma Maxi Kit workflow — sample to microvesicles to total RNA in just 1 hour). 

In the exosome purification stage, prefiltered sample (with particles larger than 0.8 μm excluded) is mixed with Buffer XBP and bound to an exoEasy membrane affinity spin column. The bound exosomes are washed with Buffer XWP, and then lysed with QIAzol. 

In the RNA extraction step, chloroform is added to the QIAzol eluate, and the aqueous phase is recovered and mixed with ethanol. Total RNA, including miRNA, binds to the spin column, where it is washed three times and eluted. 

Applications

exoRNeasy Kits are highly suited for isolating total RNA, including mRNA and miRNA, from microvesicles found in biofluid samples. 


Applications include:

  1. Transcriptomics
  2. miRNA profiling
  3. mRNA profiling
  4. RNA sequencing

Product Resources

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