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Invitrogen™ mirVana™ miRNA Isolation Kit, without phenol

Catalog No. AM1561
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AM1561 40 Preps
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Catalog No. AM1561 Supplier Invitrogen™ Supplier No. AM1561
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Description

The mirVana™ miRNA Isolation Kit uses a rapid procedure to isolate small RNAs from tissue and cells using an efficient glass fiber filter (GFF)-based method.

The mirVana™ miRNA Isolation Kit uses a rapid procedure to isolate small RNAs from tissue and cells using an efficient glass fiber filter (GFF)-based method. The method isolates total RNA ranging in size from kilobases down to 10-mers. Each kit contains sufficient reagents and consumables for either 40 isolations of total RNA (including small RNAs), or 20 separate large and small RNA fractions. Phenol must be purchased separately. Features of this kit:

• Efficient isolation of small RNA-containing total RNA
• Enrich for small RNA (less than 200 nt) to increase sensitivity in downstream analyses
• Simple 30 minute procedure
• Ideal for miRNA, siRNA, shRNA, and snRNA analysis
• Compatible with virtually all cell and tissue types

Efficient recovery of small RNA
Current RNA purification procedures rely on organic extraction, followed by alcohol precipitation or adsorption of nucleic acid molecules on a GFF or silicate matrix. Unfortunately, the former procedure requires alcohol precipitation, which is time consuming and inefficiently recovers small RNAs. The latter procedure typically results in the loss of substantial amounts of small RNA. The mirVana™ miRNA Isolation Kit uses organic extraction followed by purification on a GFF under specialized binding and wash conditions. As a result, the kit effectively recovers all RNA—from large mRNA and ribosomal RNA down to 10-mers—from virtually all cell and tissue types. The mirVana™ miRNA Isolation Kit enables the isolation of small RNA-containing total RNA from samples consisting of 102–107 cultured cells or 0.5–250 mg tissue.

Enrich for small RNA
Although the mirVana™ miRNA Isolation Kit efficiently purifies all RNA larger than 10 nt, it also includes procedures for isolating RNA fractions, specifically enriched or depleted in small RNA species. Through the enrichment procedure included in the mirVana™ miRNA Isolation Kit, RNA molecules of ∼200 nt and less can be efficiently purified away from the larger RNA species. The resulting RNA preparation is highly enriched for miRNAs, siRNAs, and/or snRNAs. Small RNA enrichment using the mirVana™ miRNA Isolation Kit procedure allows more sensitive small RNA detection with less background as compared to the same assay used with total RNA.

Specifications

Content And Storage • 30 mL miRNA Wash Solution I (room temperature)
• 50 mL Wash Solution 2/3 (room temperature)
• 80 Collection Tubes (room temperature)
• 40 Filter Cartridges (room temperature)
• 100 mL Lysis/Binding Buffer (4°C)
• 10 mL miRNA Homogenate Additive (4°C)
• 1.4 mL Gel Loading Buffer II (-20°C)
• 5 mL Elution Solution (-20°C, 4°C, or room temperature)
Isolation Technology Spin Column (Glass Fiber Filter), Organic Extraction
Purification Time 30 min. (After sample homogenization)
Elution Volume 100 μL
Sample Type Bacteria, Cells, Plant, Tissue, Viruses, Yeast, Enzymatic Reactions
Final Product Type Small RNA
High-throughput Compatibility Not High-throughput Compatible (Manual)
For Use With (Application) qRT-PCR, microarray analysis, Northern blotting
Quantity 40 Preps
Shipping Condition Room Temperature
Starting Material Amount Bacteria: ≤107 cells
Cells: ≤107
Plant: ≤250 mg
Tissue: ≤250 mg
Virus: ≤107 cells
Yeast: ≤107 cells
Enzymatic Reactions: varies
Yield Cells: ≤16 μg RNA per 50,000 cells
Tissue: ≤160 μg per 5 mg
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Frequently Asked Questions (FAQs)

What are different methods for cleaning up my miRNA sample?

- Precipitation of miRNA: Follow the instructions for the mirVANA miRNA Isolation Kit or the mirVANA PARIS RNA and Native Protein Purification Kit.
- Double phenol/guanidium (chaotropic reagent) extraction: Disrupt tissue with phenol/guanidine, then extract with phenol, centrifuge, remove the supernatant, and precipitate the RNA.
- Glass-fiber filter: Disrupt the tissue in a chaotropic agent, bind the RNA to a glass-fiber filter, then wash off unbound DNA/protein/salt/nucleotides and elute the RNA.

Can I order separately the Spin Cartridges and the Elution Tubes from the mirVana miRNA Isolation Kit, without phenol (Cat. No. AM1561)?

Yes, the Spin Cartridges and the Elution Tubes can be ordered separately. Please see the following links for the Spin Cartridges and Elution Tubes.
Do you offer a stand-alone phenol product that I can use with mirVana miRNA Isolation Kit, without phenol (Cat. No. AM1561)?

Yes, we offer Acid-Phenol: Chloroform, pH 4.5 (with IAA, 125:24:1), Cat. No. AM9720 or AM9722.
Note: We also offer mirVana miRNA Isolation Kit, with phenol (Cat. No. AM1560) that already comes with phenol.
What kit do you recommend to capture small RNA that is >17 nt in size?

We recommend using the mirVana miRNA Isolation Kit (Cat. No. AM1560, AM1561), both of which are designed to capture small RNAs, such as miRNAs, tRNAs, etc. Purification of RNAs as small as 20 nt has been obtained in-house using these kits. Note: Cat. No. AM1561 does not contain phenol, but is required for the procedure. Phenol can be purchased separately (Cat. No. AM9720).

For Research Use Only. Not for use in diagnostic procedures.