Macherey-Nagel™ NucleoSpin™ RNA Midi kit for RNA purification

NucleoSpin™ RNA L allows isolation of DNA-free high quality RNA from large amounts of starting material, e.g., cultured cells, tissue, bacteria and yeast. Cells are lysed by incubation in a solution containing large amounts of chaotropic ions. This lysis buffer immediately inactivates RNases and creates appropriate binding conditions which favour adsorption of RNA to the silica membrane. After lysis, homogenisation and reduction of viscosity are achieved by filtration with NucleoSpin™ filter L units. Contaminating DNA bound to the silica membrane is removed by a DNase I solution which is directly applied onto the silica membrane during the preparation. Optimal conditions for the DNase are achieved by washing the silica membrane with a specific desalting buffer before treatment. Salts, metabolites and macromolecular cellular components are removed by simple washing steps with two different buffers. Total RNA is finally eluted with RNase-free water. Kit components: NucleoSpin™ RNA L columns with 15mL collecting tubes, 15mL elution tubes, NucleoSpin™ filters L, buffers, RNase-free DNase I, DNase I reaction buffer, RNase-free water.

• DNase I included for removal of genomic DNA plus NucleoSpin™ filters for homogenisation and reduction of lysate viscosity
• Silica membrane technology
• Yield: 180μg (10⁷ Hela cells), 620μg (4 x 10⁷ Hela cells)
• Sample material: <5 x 10⁷ cultured cells, <10¹⁰ bacterial cells, ≤3 x 10⁸ yeast cells, <200mg tissue
• Typical RIN: >9
• Elution volume: 500μL
• Binding capacity: 700μg
• Preparation time: 80 min./4 preps
• Format: midi spin column Total RNA isolation from cultured cells, tissue, bacteria, yeast, samples stored in RNAlater and RNA clean up of reaction mixtures.