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Invitrogen™ mirVana™ miRNA Isolation Kit, with phenol
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 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.
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 10nt, 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.
• Efficient isolation of small RNA-containing total RNA
• Enrich for small RNA (less than 200nt) 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
Accessory products
For tissues that have very high levels of ribonucleases, such as pancreas or spleen, the mirVana miRNA Isolation Kit is recommended, although it is not designed to recover protein and small RNA from the same sample.
Not high-throughput compatible (Manual)
Specifications
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) • 100 mL Acid-Phenol:Chloroform (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 |
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Frequently Asked Questions (FAQs)
For tissue or cells, we recommend the mirVana miRNA Isolation Kit (Cat. No. AM1560), while for liquid samples, we recommend the mirVana Paris RNA and Native Protein Purification Kit (Cat. No. AM1556). For high-throughput miRNA isolation we recommend using the MagMAX mirVana Total RNA Isolation kit (Cat. No. A27828)
- 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.
One to ten nanograms of total RNA per 15 µL RT reaction is recommended for use with the TaqMan MicroRNA Assay. We have successfully detected miRNAs in as little as 25 pg of total RNA (~100 cells) in samples where the miRNA was highly abundant. The total RNA isolation procedure used must preserve small RNA species. We recommend the Invitrogen mirVana miRNA Isolation Kit (Cat. No. AM1560) or TRI reagent (Cat. No. AM9738).
Yes, the RNA lysates can be stored at -80 degrees C for up to 2 months.
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.