NucleoSpin® miRNA

Catalog No. 740971

£1

Parallel isolation of small and large RNA

Available: In stock

Product Description

Small and large RNA isolation
No toxic phenol / chloroform convenient extraction
Parallel isolation of small and large RNA

• RNA purification fractionated by size :
– Isolation of small RNA only (< 200 b),
– Isolation of small RNA (< 200 b) and large RNA (> 200 b) in two separate fractions
– Isolation of total RNA (small and large RNA in one fraction)

• Additional isolation of total protein fraction ready to use for SDS-PAGE and Western blot analysis

• Excellent RNA recovery and purity by chaotropic salt lysis without phenol/chloroform (patent pending)

• NucleoSpin® Filters for efficient sample homogenization

• rDNase for efficient on-column removal of genomic DNA

• Silica-membrane technology

• Sample material: < 107 cultured cells, < 30 mg human / animal tissue, < 50 mg plant tissue, < 150 µL reaction mixture

• Typical yield: 10 µg small RNA, 95 µg large RNA (107 HeLa cells)

  • Kit contents :
    • NucleoSpin® RNA Columns, NucleoSpin® miRNA Columns, NucleoSpin® Protein Removal Columns, Collection Tubes (2 mL, 2 mL lid, 1.5 mL), NucleoSpin® Filters, buffers, RNase-free rDNase
  • Reference numbers :
    • 10 preps: 740971.10
    • 50 preps: 740971.50
    • 250 preps: 740971.250
  • Parallel isolation of small and large RNA from human / animal tissue and cultured cells, from plant tissue  and in combination with phenol / chloroform (e.g., TRIzol®) lysis
  • Purification of siRNA and large dsRNA from DICER reactions
  • Typical downstream applications: real-time RT-PCR, Northern blotting, chip hybridization

NucleoSpin® miRNA is suitable for simultaneous isolation of small RNA (< 200 nt, e.g., miRNA, pre-miRNA, tRNA, 5S RNA), large RNA (> 200 nt, e.g., mRNA, 18S rRNA, 28S rRNA, pri-miRNA), and protein in three separate fractions from a large variety of sample materials :

  1. Mechanical disruption of the sample material in Lysis Buffer ML.
  2. Convenient homogenization and clearing of crude lysates with NucleoSpin® Filters (violet rings).
  3. Addition of ethanol to adjust binding conditions for DNA and large RNA.
  4. Binding of DNA and large RNA to the NucleoSpin® RNA Column (blue ring). The flow-through of the NucleoSpin® RNA Column contains small RNA and protein.
  5. Removal of residual genomic DNA by on-column digestion with the provided RNase-free recombinant DNase.
  6. Washing of large RNA and elution with RNase-free water → large RNA fraction.
  7. Addition of Protein Precipitation Buffer MP to the flow-through of the NucleoSpin® RNA Column (step 4) to precipitate the protein.
  8. Collection of protein precipitate by centrifugation → protein fraction. Filtration of the supernatant through a NucleoSpin® Protein Removal Column (white ring) to completely remove the residual protein precipitate to achieve best possible purity of the small RNA fraction. The flow-through of the NucleoSpin® Protein Removal Column only contains small RNA.
  9. Addition of Binding Buffer MX to adjust binding conditions for small RNA.
  10. Binding of small RNA to the NucleoSpin® miRNA Column (green ring).
  11. Washing of small RNA and elution with RNase-free water → small RNA fraction

The precipitated protein can easily be dissolved in Laemmli buffer and used for SDS-PAGE, Western Blot analysis, and protein quantification, for example with the MACHEREY-NAGEL Protein Quantification Assay.
The eluted RNA and miRNA are ready-to-use for all standard downstream applications, for example RT-PCR, Northern Blot, or chip hybridization.

  • Eur J Hum Genet. 2015 Aug;23(8):1100-5. doi: 10.1038/ejhg.2014.244. Epub 2014 Nov 12.
    Expression analysis in intestinal mucosa reveals complex relations among genes under the association peaks in celiac disease.
    Plaza-Izurieta L, Fernandez-Jimenez N, Irastorza I
    View Abstract

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