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miRNeasy Serum/Plasma Kit

For purification of cell-free total RNA, including miRNA, from animal and human plasma and serum

Features

  • Internal control available for normalization
  • Minimal elution volume
  • High-purity RNA suitable for all downstream applications
  • Easy, robust procedures
  • Automatable protocol
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miRNeasy Serum/Plasma Kit (50)

Cat. No. / ID: 217184

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
HK$4,770.00
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The miRNeasy Serum/Plasma Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

Product Details

The miRNeasy Serum/Plasma Kit is designed for purification of cell-free total RNA — primarily miRNA and other small RNA — from small volumes of serum and plasma. RNA from serum and plasma typically consists of molecules <100 nucleotides. Purification can be automated on the QIAcube Connect.

 

The NEW miRNeasy Serum/Plasma Advanced Kit – get excellent results faster without phenol/chloroform using our QIAzol-free Advanced chemistry. Try it today.

Performance

The miRNeasy Serum/Plasma Kit efficiently purifies RNA from up to 200 µl serum or plasma. RNA prepared using miRNeasy Kits is highly pure and ready for use in sensitive downstream applications. miRNeasy procedures minimize the possibility of contamination with salts or phenol, which could interfere with later analyses.

Principle

One of the most exciting areas of current miRNA research involves the assessment of miRNAs present in serum or plasma samples. The presence of relatively stable, extracellular miRNAs in serum and plasma has generated great interest in the potential use of changes in these miRNA levels as noninvasive biomarkers for a variety of diseases. Using RNA purified from serum or plasma, unique expression of several miRNAs has been shown for some cancers. The miRNeasy Serum/Plasma Kit enables RNA purification from small volumes of plasma or serum. RNA from serum and plasma typically consists of molecules <100 nucleotides. Purified RNA can then be used for biomarker discovery.

The miRNeasy Serum/Plasma Kit combines phenol/guanidine-based lysis of samples and silica-membrane-based purification of total RNA. QIAzol Lysis Reagent, included in the kit, 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.

Procedure

Serum or plasma samples are lyzed in QIAzol Lysis Reagent. After addition of chloroform, the lysate is separated into aqueous and organic phases by centrifugation. RNA partitions to the upper, aqueous phase, while DNA partitions to the interphase and proteins to the lower, organic phase or the interphase.

The upper, aqueous phase is extracted, and ethanol is added to provide appropriate binding conditions for all RNA molecules from approximately 18 nucleotides upwards. The sample is then applied to the RNeasy MinElute spin column, where the total RNA binds to the membrane and phenol and other contaminants are efficiently washed away. High-quality RNA is then eluted in a small volume of RNase-free water.

Serum and plasma contain primarily small RNAs, therefore separate purification of small and large RNA fractions is not necessary.

Automated miRNA purification on the QIAcube

Purification of total RNA, including miRNA, from the aqueous phase after phase separation can be automated on the QIAcube. The innovative QIAcube 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 follows the same steps as the manual procedure (i.e., lyse, bind, wash, and elute), enabling you to continue using the miRNeasy Serum/Plasma Kit for purification of high-quality RNA.

Applications

The miRNeasy Serum/Plasma Kit allows purification of cell-free total RNA, including miRNA, for use in a variety of applications, including:

  • Biomarker discovery
  • miRNA profiling
  • miRNA sequencing
  • miRNA analysis
  • Northern blot analysis
  • Microarray analysis

FAQ

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 is the Spike-in-Control in the miRNeasy Serum/Plasma Kit ?

The miRNeasy Serum/Plasma Spike-In Control (219610) is ordered separately from the miRNeasy Serum/Plasma Kit. It is 10 pmol lyophilized C. elegans miR-39 miRNA mimic.

FAQ ID -3169
What is the bacterial ribosomal RNA recommended to use in the miRNeasy Serum/Plasma Protocol?

It is bacterial ribosomal RNA (Roche), cat. no. 10 206 938 001.

FAQ ID -3173
Do you have a protocol for the isolation of genomic DNA and/or proteins from fatty tissue treated with QIAzol?
FAQ ID -955
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
3339 - Does the miRNeasy Serum/Plasma Spike-in Control (cat n° 219610) negatively impact subsequent NGS sequencing?

The miRNeasy Serum/Plasma Spike-in Control (cat n° 219610)  will not pose a problem for NGS sequencing. In case of a library prep for larger RNAs, a size selection step may be part of the procedure that removes small RNA in that size range.

How do I prepare the Spike-in-Control catalog # 219610 to get copies/μl?

Reconstitute by adding 300 μl RNase-free water per tube, resulting in a 2 x 10^10 copies/μl stock. miRNeasy Serum/Plasma Spike-In Control stock should be stored at –80°C. For large volumes, first aliquot into smaller volumes prior to long-term storage at –80°C.

Refer to page 25 of the miRNeasy Serum/Plasma Protocol Handbook for Preparation of miRNeasy Serum/Plasma Spike-In Control

http://www.qiagen.com/products/catalog/sample-technologies/rna-sample-technologies/mirna/mirneasy-serumplasma-kit#resources

FAQ ID -3170
What is the miRNA yield from the miRNeasy Serum/Plasma Kit?

Yields of total RNA achieved with the miRNeasy Serum/Plasma Kit vary strongly between samples from different individuals. However, yield of RNA from 50-200 µl human plasma and serum is not quantifiable with photometric methods like NanoDrop; they are usually too low for quantification by OD measurement.

You may use more sensitive colorimetric or fluorimetric assays for quantification, i.e. RiboGreen assays.

Use of miRNeasy Serum/Plasma Spike-In Control (cat. no. 219610) and corresponding Ce_miR-39_1 miScript Primer Assay (cat.no. MS00019789) is recommended to monitor miRNA purification and amplification.

FAQ ID -3171
3338 - Does the miRNeasy Serum/Plasma Spike-in Control (cat n° 219610) have 3'-hydroxyl and 5'-phosphate groups?

Both ends of the miRNeasy Serum/Plasma Spike-in Control carry only the hydroxyl groups. The 5'-phosphate is not relevant for the biological function.