microRNA Purification Kit

In stock
SKU
1021300
210,00 €
isolation of small RNAs resulting in minimal contamination of larger RNA and gDNA. Isolate very rapidly miRNA, siRNA, tRNA and 5S rRNA without the use of phenol or chloroform.Purified miRNA is suitable for microarray analysis.
Supplier
Norgen Biotek
Cat. no.
1021300
Kit size
25
Links
Norgen Biotek
Protocol
Information sheet

Features

  • Rapid spin-column format allows for the processing of 10 samples in 30 minutes.
  • RNA is isolated without the use of harmful chemicals such as phenol or chloroform.
  • All small RNA species can be isolated including miRNA, siRNA, tRNA and 5S rRNA.
  • Efficient isolation of small RNA species using a 2 column process, resulting in minimal contamination of larger RNA and genomic DNA.
  • Purified RNA can be used in a number of downstream applications relating to gene regulation and functional analysis, including northern blotting and microarray analysis.

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, bacterial cells, plants and blood. These small RNAs include regulatory RNA molecules such as microRNA (miRNA) and short interfering RNA (siRNA), as well as tRNA and 5S rRNA. 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 RT-PCR, northern blotting and microarray analysis.
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 without the use of phenol or chloroform. The process involves the use of two different spin columns: the Large RNA Removal Column and the microRNA Enrichment Column (please see flow chart on page 3). Briefly, the cells or tissues of interest are lysed using the provided Lysis Buffer, and ethanol is then added to the sample. The lysate is then applied to the Large RNA Removal Column, and 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 microRNA Enrichment Column. The small RNA molecules will then bind to the resin, and any impurities are removed through a series of 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.
Column Binding Capacity 50 µg
Maximum Column Loading Volume 600 µl
Size of RNA purified <200 nt
Maximum Amount of Starting Material: Animal Cells: 3x10exp6 cells Animal Tissues: 5-25 mg Bacteria: 1x10exp9 cells Plant Tissues: 50 mg Blood: 100 µl
Time to Complete 10 Purifications 25 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.


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