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miRNeasy Micro Kit

For purification of miRNA and total RNA from small amounts of cells and tissues

  • Effective purification of miRNA and total RNA from small samples
  • High-purity RNA suitable for all downstream applications
  • Easy, robust procedures
  • Automatable protocols

The miRNeasy Micro Kit is designed for purification of total RNA, including miRNA and other small RNA molecules, from small amounts of cultured cells, sorted cells, laser capture microdissection (LCM) samples from cryosections, and various animal and human tissues. Purification can be automated on the QIAcube.

Cat No./ID: 217084
miRNeasy Micro Kit (50)
€375.00
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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
The miRNeasy Micro Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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High, reproducible recovery of miRNA from small cell numbers.

Total RNA was purified from 10-fold serial dilutions of Jurkat cells (106 to 10 cells) using the miRNeasy Micro Kit. RNA was purified from 2 replicates of each dilution, either [A] manually or [B] using the QIAcube. Purified RNA was used to detect miR-16 using the miScript PCR System on a Rotor-Gene Q cycler. miR-16 was detected in as few as 10 cells and CT values were consistently reproducible. No signal was detected in negative controls (blue and yellow curves).

Performance
The miRNeasy Micro Kit efficiently purifies RNA from small tissue and cell 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.

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. In addition to higher purity, the miRNeasy Micro Kit provides superior yields to alternative methods of miRNA purification, such as using phenol-based extraction and precipitation.
Principle

The miRNeasy Micro Kit is designed for purification of total RNA, including miRNA and other small RNA molecules, from small amounts of cultured cells (up to 1 x 106 cells) and various animal and human tissues (up to 5 mg tissue). The 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. After sample homogenization in QIAzol Lysis Reagent, chloroform is added and following centrifugation, RNA partitions to the upper, aqueous phase. The aqueous phase is extracted and RNA is purified on the RNeasy MinElute spin column, while contaminants are washed away. The unique design of RNeasy MinElute spin columns enables minimal elution volumes, as little as 10 µl, to concentrate the RNA.

Procedure

The miRNeasy Micro Kit enables 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.

Automated miRNA purification on the QIAcube

Purification of total RNA, including miRNA, from the aqueous phase after phase separation can be fully automated on the QIAcube (see figure "High, reproducible recovery of miRNA from small cell numbers"). 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 Micro Kit for purification of high-quality RNA.

Applications

The miRNeasy Micro Kit allows purification of miRNA with total RNA for use in a variety of applications, including:

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

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Kit Handbooks (1)
For purification of total RNA, including miRNA, from animal and human cells and tissues
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Safety Data Sheets (1)
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