High Pure miRNA Isolation Kit
Low to medium throughput miRNA isolation.
For life science research only. Not for use in diagnostic procedures.
The High Pure miRNA Isolation Kit purifies and enriches small RNAs, such as microRNA (miRNA) from animal cells and tissue samples (including formalin-fixed, paraffin-embedded sections) or plant material. It can also be used to purify total RNA or to prepare samples enriched for small RNAs (<100 nucleotides).
- Isolate DNA-free miRNA ideal for qualitative and quantitative reverse transcription. ideal for qualitative and quantitative reverse transcription.
- Obtain excellent performance and linearity in RT-PCR.
- Generate stable , highly pure miRNAs using a simple, efficient protocol.
- Purify miRNA without using hazardous organic solvents. Avoid phenol/chloroform steps required by other suppliers' miRNA purification kits.
- Choose one flexible kit for all your miRNA purifications. Use the same kit to purify small RNAs from a variety of sample types (see Figure 1).
- Tissue Lysis Buffer (for FFPE sections)
- Proteinase K, recombinant (for FFPE sections)
- Binding Buffer
- Binding Enhancer
- Wash Buffer
- Elution Buffer
- High Pure Filter Tubes
- Collection Tubes
The High Pure miRNA Isolation kit rapidly purifies RNA that is suitable for direct use in many downstream applications:
- Northern blotting
- cDNA synthesis
- Primer extension
- miRNA array hybridization
- Relative quantification of miRNA with RT-PCR using, for example, the LightCycler® 480 System
Figure 1: Comparison of miRNA yields from different tissues using the High Pure miRNA Isolation Kit versus kits from other suppliers.
MicroRNAs are natural regulators of gene expression, affecting almost every cellular process (growth and development, cell differentiation, cell death). They occur naturally in diverse animal and plant species, and hundreds have been discovered since the late 1990s. A single miRNA can ramp down expression of multiple genes. Nucleic acids bind to the surface of the glass fiber fleece in the presence of a chaotropic salt (guanidine HCl). This allows the High Pure filter tube to specifically immobilize nucleic acids (both DNA and RNA) while they are freed of contaminants.
Capacity: The High Pure Spin Filter Tubes hold up to 700 μL sample volume.
Typical miRNA Recovery
Starting Material and Quantity
|Yield/ Recovery||Time Required||Number of Reactions|
Animal tissue, animal cell culture, formalin-fixed, paraffin-embedded tissue
Depending on miRNA
In the presence of a chaotropic salt, RNA binds selectively to special glass fibers prepacked in the High Pure Spin Filter Tube. Bound RNA is purified in a series of rapid wash-and-spin steps to remove contaminating salts, proteins, and other cellular impurities, and then eluted using a low-salt solution. By lowering the concentration of Binding Enhancer during the binding step in the two-column protocol, the small RNA-containing miRNA passes the first column unbound. When the concentration of Binding Enhancer is increased, the small RNA fraction can be bound to a second High Pure Spin Filter Tube.
More than 60% recovery is obtained when 1 μg miRNA is added to106 K-562 cells and isolated with the one-column method, and more than 50% is recovered with the two-column method.