05080576001

High Pure miRNA Isolation Kit

Properties

• Quality Level: 100
• mfr. no.: Roche
• packaging: kit of for 50 isolations

Description

General description

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.

High Pure miRNA Isolation Kit; Instructions For Use

Low to medium throughput miRNA isolation.

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).

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.

Application

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

Features and Benefits

• 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.

Components

• 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

Quality

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.

Other Notes

For life science research only. Not for use in diagnostic procedures.

Relative Quantification of miRNA Target mRNAs by Real-Time qPCR

Cancer Research - Gene Expression Analysis You Can Count On Article (PDF, 622 KB)

High Pure miRNA Isolation Kit Article (PDF, 176.3 KB)

High Pure Technology and Silica Adsorption Kits

Legal Information

LightCycler is a registered trademark of Roche

Safety Information

• Pictograms: GHS05,GHS07,GHS08
• Signal Word: Danger
• Hazard Statements: H302 + H312 + H332 - H314 - H317 - H334 - H335 - H360Df - H412
• Precautionary Statements: P201 - P260 - P280 - P303 + P361 + P353 - P304 + P340 + P310 - P305 + P351 + P338 + P310 - P308 + P313
• Target Organs: Respiratory system
• RIDADR: NONH for all modes of transport
• WGK Germany: WGK 2
• Flash Point(F): 285.8 °F
• Flash Point(C): 141 °C