exoRNeasy Midi and Maxi Kits

For efficient isolation of exosomal RNA from biofluids

S_1084_5_GEN_V2

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exoRNeasy Midi Kit (50)

Cat. No. / ID:  77144

For purification of total exosome-derived RNA; includes 50 exoEasy Midi columns and 50 RNeasy MinElute spin columns
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€1,591.00
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KitReagentBuffer
exoRNeasy Kit
Reagent UI
Buffer
Column type
Midi
Maxi
Midi/Maxi
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.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

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

Product Details

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.

Want to try this solution for the first time? Request a trial kit quote.

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).

See figures

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.

See figures

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

Supporting data and figures

Publications

Characterization of RNA from Exosomes and Other Extracellular Vesicles Isolated by a Novel Spin Column-Based Method.
Enderle D; Spiel A; Coticchia CM; Berghoff E; Mueller R; Schlumpberger M; Sprenger-Haussels M; Shaffer JM; Lader E; Skog J; Noerholm M;
PLoS One; 2015; 10 (8):e0136133 2015 Aug 28 PMID:26317354
miRNA contents of cerebrospinal fluid extracellular vesicles in glioblastoma patients.
Akers JC; Ramakrishnan V; Kim R; Phillips S; Kaimal V; Mao Y; Hua W; Yang I; Fu CC; Nolan J; Nakano I; Yang Y; Beaulieu M; Carter BS; Chen CC;
J Neurooncol; 2015; 123 (2):205-16 2015 Apr 23 PMID:25903655
Urinary Exosomal miRNA Signature in Type II Diabetic Nephropathy Patients.
Delić D; Eisele C; Schmid R; Baum P; Wiech F; Gerl M; Zimdahl H; Pullen SS; Urquhart R;
PLoS One; 2016; 11 (3):e0150154 2016 Mar 1 PMID:26930277
Curcumin inhibits in vitro and in vivo chronic myelogenous leukemia cells growth: a possible role for exosomal disposal of miR-21.
Taverna S; Giallombardo M; Pucci M; Flugy A; Manno M; Raccosta S; Rolfo C; De Leo G; Alessandro R;
Oncotarget; 2015; 6 (26):21918-33 2015 Sep 8 PMID:26116834
The Swine Plasma Metabolome Chronicles "Many Days" Biological Timing and Functions Linked to Growth.
Bromage TG; Idaghdour Y; Lacruz RS; Crenshaw TD; Ovsiy O; Rotter B; Hoffmeier K; Schrenk F;
PLoS One; 2016; 11 (1):e0145919 2016 Jan 6 PMID:26735517