miRNeasy Kits

For purification of miRNA and total RNA from tissues and cells

Features

  • Effective microRNA (miRNA) and total RNA purification, even from small samples
  • Efficient enrichment of miRNA and RNAs <200 nucleotides
  • Protocols for copurification or isolation of separate fractions
  • High-purity RNA suitable for all downstream applications
  • Automatable protocols and high-throughput processing in 96-well format
miRNeasy Micro Kit (50)

Cat. No. / ID: 217084

For 50 total RNA preps: 50 RNeasy MinElute Spin Columns, Collection Tubes (1.5 ml and 2 ml), QIAzol Lysis Reagent, RNase-Free Reagents and Buffers
€426.00
Column typePlate type
Micro
Mini
96 well
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miRNeasy Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.

Product Details

miRNeasy Kits enable purification of total RNA from all types of animal tissues and cells, including difficult-to-lyse tissues and small tissue and cell samples. Alternatively, an miRNA-enriched fraction and a total RNA (>200 nt) fraction can be purified separately (for separate purification, an RNeasy MinElute Cleanup Kit is required when using the miRNeasy Micro or Mini Kit and an additional RNeasy 96 plate when using the miRNeasy 96 Kit). Purification of 1–12 samples can be automated on the QIAcube Connect. High-throughput processing in convenient 96-well format can be performed using centrifugation or vacuum procedures.

The NEW miRNeasy Tissue/Cells Advanced Mini Kit – get excellent results faster without phenol/chloroform using our QIAzol free Advanced chemistry. Try it today.

Performance

QIAzol based microRNA purification (miRNA purification) using miRNeasy Kits efficiently purifies RNA from small tissue (up to 5 mg tissue) and cell (up to 1 x 106 cells) samples (see figure " High, reproducible recovery of miRNA from small cell numbers"). RNA can be purified from a variety of tissues and cells, including difficult-to-lyse tissues.

Purification of total RNA including miRNA allows direct comparison of miRNA expression levels with those of housekeeping reference genes or any other mRNA of interest. Alternatively, for some sensitive downstream applications, an miRNA-enriched fraction without larger RNA may be required. miRNeasy Kits enable efficient enrichment of RNA down to approximately 18 nt in size. The presence of very small RNAs is clearly visible after purification using an miRNeasy Kit, in contrast to samples prepared using an alternative kit (see figure " Effective enrichment of small RNA"). miRNeasy Kits efficiently purify RNA from both tissues and cells, even when low amounts of starting material are used (see figure " Effective purification from a range of starting amounts").

RNA can be purified with miRNeasy Kits from a variety of tissues and cells, including difficult-to-lyse tissues (see figure " Efficient copurification from a wide range of tissues"). miRNeasy procedures eliminate the possibility of contamination with salts or phenol which could interfere with later analyses (see figure " Highly pure RNA without phenol carryover"). In addition to higher purity, miRNeasy Kits offer superior yields to alternative methods of miRNA purification, such as using TRIzol Reagent (see figure " miRNeasy Mini Kit outperforms TRIzol").

RNA prepared using miRNeasy Kits is highly pure and ready for use in sensitive downstream applications.

See figures

Principle

miRNeasy Kits combine phenol/guanidine-based lysis of samples with silica membrane-based purification of total RNA. QIAzol Lysis Reagent, included in the kits, is a monophasic solution of phenol and guanidine thiocyanate, designed to facilitate lysis of tissues, to inhibit RNases, and also to remove most of the cellular DNA and proteins from the lysate by organic extraction.

miRNeasy Micro and Mini Kits are automatable on the QIAcube Connect. The miRNeasy 96 Kit enables vacuum and/or spin processing.

Procedure

miRNeasy Kits enable purification of total RNA, which includes RNA from approximately 18 nucleotides (nt) upwards. Alternatively, an miRNA-enriched fraction and a total RNA (>200 nt) fraction can be purified separately (see flowchart " miRNeasy procedures"). Total RNA, including miRNA, is highly suitable for use in miRNA quantification by real-time RT-PCR. Enrichment of small RNAs in a separate fraction may be advantageous for certain applications where mRNA and rRNA could lead to increased background. This specialized protocol requires use of the RNeasy MinElute Cleanup Kit with the miRNeasy Micro or Mini Kit. If using the miRNeasy 96 Kit, 2 RNeasy 96 plates are required for every 96 samples.

Automated miRNA purification on the QIAcube Connect

Purification of total RNA, including miRNA, from the aqueous phase after phase separation can be fully automated on the QIAcube Connect using the miRNeasy Micro or Mini Kit (see figure " High, reproducible recovery of miRNA from small cell numbers"). The innovative QIAcube Connect uses advanced technology to process QIAGEN spin columns, enabling seamless integration of automated, low-throughput sample prep into your laboratory workflow. Sample preparation using the QIAcube Connect follows the same steps as the manual procedure (i.e., lyse, bind, wash, and elute), enabling you to continue using the miRNeasy Micro or Mini Kit for purification of high-quality RNA.

See figures

Applications

miRNeasy Kits allow purification of miRNA with total RNA for use in a variety of applications, including:

  • RNA-seq
  • Quantitative, real-time RT-PCR (e.g., using the miScript PCR System)
  • Northern blot analysis
  • Microarray analysis

Comparison of miRNeasy Kits

Features miRNeasy Micro Kit miRNeasy Mini Kit miRNeasy 96 Kit
Applications PCR, qPCR, real-time RT-PCR,
microarray
PCR, qPCR, real-time RT-PCR,
microarray
PCR, qPCR, real-time RT-PCR,
microarray
Elution volume 10 µl 30 µl Min. 2 x 45 µl
Format MinElute Spin column Spin column 96-well plate
Main sample type Tissue, cells Tissue, cells Tissue, cells
Processing Manual (centrifugation or vacuum) Manual (centrifugation or vacuum) Manual (centrifugation or vacuum)
Purification of total RNA, miRNA,
poly A+ mRNA, DNA or protein
miRNA, total RNA miRNA, total RNA miRNA, total RNA
Sample amount 5 mg (10 mg for adipose) tissue,
1 x 106 cells
50 mg (100 mg for adipose) tissue,
1 x 107 cells
50 mg (100 mg for adipose) tissue,
1 x 107 cells
Technology Silica technology Silica technology Silica technology
Time per run or per prep <2 hours
Yield Varies Varies Varies

Supporting data and figures

FAQ

How much RNA does a typical mammalian cell contain?

The RNA content and RNA make up of a cell depend very much on its developmental stage and the type of cell. To estimate the approximate yield of RNA that can be expected from your starting material, we usually calculate that a typical mammalian cell contains 10–30 pg total RNA.

 

The majority of RNA molecules are tRNAs and rRNAs. mRNA accounts for only 1–5% of the total cellular RNA although the actual amount depends on the cell type and physiological state. Approximately 360,000 mRNA molecules are present in a single mammalian cell, made up of approximately 12,000 different transcripts with a typical length of around 2 kb. Some mRNAs comprise 3% of the mRNA pool whereas others account for less than 0.1%. These rare or low-abundance mRNAs may have a copy number of only 5–15 molecules per cell.

FAQ ID -2946
I received a kit containing the MinElute columns; however, they were left out for a while and not stored at 2–8°C upon receipt. Can I still use them?

The MinElute spin columns included in the following kits should be stored at 2–8°C upon arrival: AllPrep DNA/RNA Micro, EpiTect Fast DNA Bisulfite, EpiTect Fast FFPE Bisulfite, EpiTect Fast LyseAll Bisulfite, EpiTect Plus DNA Bisulfite, EpiTect Plus FFPE Bisulfite, EpiTect Plus LyseAll Bisulfite, exoRNeasy Serum/plasma Maxi, exoRNeasy Serum/Plasma Midi, GeneRead DNA FFPE, GeneRead rRNA Depletion, GeneRead Size Selection, MinElute Gel Extraction, MinElute PCR Purification, MinElute Reaction Cleanup, miRNeasy FFPE, miRNeasy Micro, miRNeasy Serum/Plasma, QIAamp DNA FFPE, QIAamp DNA Investigator, QIAamp DNA Micro, QIAamp MinElute Media, QIAamp MinElute Virus Spin, QIAamp MinElute Virus Vacuum, RNeasy FFPE, RNeasy Micro, RNeasy Plus Micro.

Short-term storage (up to 4 weeks) at room temperature (15–25°C) does not affect the performance. However, for optimal performance and quality, storage temperature should not exceed 25°C.

FAQ ID - 3560
What can I use to isolate RNA smaller than 200 nucleotides?

For the isolation of microRNA (miRNA) specifically, we have developed the miRNeasy Mini Kit and the miRNeasy 96 Kit for isolation from cells and tissues.   We also have the PAXgene Blood miRNA and Tissue miRNA kits for isolation from blood stored in PAXgene Blood RNA tubes and PAXgene Tissue Containers, respectively.  Other miRNA isolation supplementary protocols can also be found by searching our comprehensive protocols at http://www.qiagen.com/literature/default.aspx?WT.svl=m.

FAQ ID -115
Do you have a protocol for the isolation of genomic DNA and/or proteins from fatty tissue treated with QIAzol?
FAQ ID -955
What is the composition of Buffer RWT?
The exact composition of Buffer RWT is confidential. Buffer RWT is a proprietary component of, for example, the miRNeasy Mini Kit and the RNeasy Plus Universal Kit. Guanidine salt and ethanol are important ingredients in Buffer RWT. Ethanol is added by the user prior to the first use of the kit. Buffer RWT is a stringent washing buffer used after preclearing the sample with QIAzol Lysis Reagent, especially if isolation of small RNAs, for example, microRNAs or RNAs from formalin-fixed tissue, is desired
FAQ ID -2798
How do I clean up RNA preparations containing miRNA?

RNA preparations containing miRNA can be cleaned up by modifying* the cleanup protocols listed in the handbooks of the RNeasy Mini Kit or the RNeasy MinElute Cleanup Kit .

 

* Modify the cleanup protocol at step 2, by increasing the volume of ethanol (96-100%) from 250 µl to 950 µl.

FAQ ID -3002
Can small miRNA-containing RNA fractions be separated from large RNAs using the miRNeasy FFPE Kit?

Unlike the Appendix A Protocol for the miRNeasy Mini Kit, which allows removal of larger RNAs such as mRNA and rRNA to enrich miRNA in a separate small RNA fraction, this is not practical using the miRNeasy FFPE Kit. RNA from FFPE- or other compromised materials is usually strongly fragmented already. Significant amounts of rRNA and mRNA fragments would end up in the small RNA-enriched fraction.

 

FAQ ID -1737
What is the composition of Buffer RPE?
The exact composition of Buffer RPE is confidential. Buffer RPE is a mild washing buffer, and a proprietary component of RNeasy Kits. Its main function is to remove traces of salts, which are still on the column due to buffers used earlier in the protocol. Ethanol, which is added by the user just before using the kit for the first time, is an important ingredient of Buffer RPE.
FAQ ID -2797
Can I use total RNA for the miRNA PCR Arrays or Assays?
Yes, you can. In fact, total RNA is the recommended starting material for the miScript PCR System. We recommend using the miRNeasy Mini Kit (217004) to isolate total RNA for use with the miScript PCR System.
FAQ ID -2726