microRNA Purification Kit

User Manual
Specifications
Product Category: microRNA Purification
Sample Type: Mammalian Cells, Tissues & More

Product Description

  • Fast and easy processing using rapid spin-column format
  • No phenol or chloroform extractions
  • Isolate all small RNA molecules (<200 nt)
  • Minimal contamination from large RNA molecules and genomic DNA
  • High quality small RNA can be used in various downstream applications

Norgen's microRNA Purification Kit provides a rapid method for the isolation and purification of small RNA molecules (< 200 nt) from cultured animal cells, small tissue samples, and bacterial cells. These small RNAs include regulatory RNA molecules such as microRNA (miRNA) and short interfering RNA (siRNA), as well as tRNA, 5S rRNA, and 5.8SrRNA. Small RNA molecules are often studied due to their ability to regulate gene expression. miRNAs and siRNAs are typically 20-25 nucleotides long, and regulate gene expression by binding to mRNA molecules and affecting their stability or translation. The small RNA molecules isolated using Norgen's microRNA Purification Kit can be used in various downstream applications relating to gene regulation and functional analysis, including northern blotting and microarray analysis.

Norgen's Purification Technology

Purification is based on spin column chromatography using Norgen's proprietary resin as the separation matrix. Norgen's resin binds RNA in a manner that depends on ionic concentrations.

The small RNA molecules are preferentially purified from other cellular components such as ribosomal RNA and proteins, without the use of phenol or chloroform. The process involves the use of two separate spin columns. Briefly, the cells or tissues of interest are lysed using the provided Lysis Buffer. Ethanol is then added to the sample, and after centrifugation the cleared lysate is applied to the first column. The larger RNA molecules will bind to the resin in the spin column, while the smaller RNA species will pass through into the flowthrough. Ethanol is then added to the flowthrough, and the sample is applied to the second spin column. The small RNA molecules will then bind to the resin, and impurities are removed through a series of two washes with the provided Wash Solution. The small RNA molecules are then eluted using the Elution Buffer, and are ready for use in various applications.

Kit Specifications

- Column binding capacity: 50 µg

- Maximum column loading volume: 700 µL

- Size of RNA purified: < 200 nt

- Maximum amount of starting material: Animal Cells: 1 x 106 cells; Animal Tissues: 2 mg; Bacteria: 1 x 109 cells

- Time to complete 10 purifications: 45 minutes

HeLa microRNA. Small RNA was isolated from HeLa cells using Norgen's microRNA Purification Kit (lanes C and D) and a competitor's kit (lanes A and B). Samples of the purified small RNA were run on an 8% urea-agarose gel. Lane M is Norgen's RNA ladder. Note that Norgen's kit is isolating only the small (<200 nt) RNA species, with no contaminating larger RNA.

Efficient fractionation of the large from the small RNA species. Norgen's microRNA Purification Kit was used to separate HeLa cell small RNA from the large RNA species. Essentially, total HeLa cell RNA is passed in the column under conditions that bind the large molecular weight RNA but not the small RNA. Samples were run on a formaldehyde-agarose gel to visualize the larger RNA species that are being removed. Lane M is a marker, Lanes A and B contain the large RNA and Lanes C and D contain the small RNA species.

Product Citations

  • Catalog Number
    21300-NB
  • Supplier
    Norgen Biotek
  • Size
  • Shipping
    RT
  • Price
  • 294,00 €
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