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Invitrogen™ RiboPure™ RNA Purification Kit
Description
The Ambion™ RiboPure™ Kit is designed for rapid purification of high-quality RNA from tissue samples or cultured cells. The kit combines TRI Reagent™ with glass fiber filter purification to yield exceptionally pure RNA, free of residual proteins and lipids. The kit includes sufficient reagents for 50 purifications from 1–100 mg of tissue or 0.1–20 x 106 cultured cells each.
• Yields the highest purity RNA attainable
• Ideal for the most stringent downstream applications
• Handles problematic samples with ease
• Fast (less than 30 minutes) and easy procedure
RNA isolated using the RiboPure™ Kit is suitable for use in all common downstream applications, including cDNA synthesis, real-time and end-point RT-PCR, microarray analysis, northern blots, and RNase protection assays. The RiboPure™ protocol is especially suited for processing samples that are difficult to disrupt, or samples that have large amounts of ribonuclease or other contaminants. Samples are first homogenized in TRI Reagent™, a monophasic solution containing phenol and guanidine thiocyanate, to rapidly lyse cells and inactivate nucleases. Addition of bromochloropropane (BCP) results in separation of the homogenate into aqueous and organic phases. RNA partitions to the aqueous phase, while DNA and protein remain in the interphase and organic phase. The aqueous phase is removed and passed through a glass fiber filter that binds the RNA. The filter is washed, and the RNA is eluted. The procedure is compatible with tissues that have been stored in Ambion's RNAlater™. Total RNA yield is typically 100–500 μg per 100 mg of tissue, depending on the type of tissue.
Order Info
Shipping Condition: Room temperature
Specifications
Specifications
| Content And Storage | • 50 mL TRI Reagent; store at 4°C • 50 Filter Cartridges; room temperature • 100 Collection Tubes (2 mL); room temperature • 60 mL Wash Solution (concentrated); room temperature • 11 mL Elution Buffer; room temperature |
| Isolation Technology | Spin Column (Glass Fiber Filter) |
| Purification Time | 30 min. (After sample homogenization) |
| Elution Volume | 100 μL |
| Sample Type | Cells, Tissue |
| Final Product Type | Total RNA |
| High-throughput Compatibility | Not High-throughput Compatible (Manual) |
| For Use With (Application) | RT-PCR, qPCR, cDNA Library Construction, NGS, Microarray Analysis, Blot Hybridization, Northern Blotting, In Vitro Translation, Nuclease Protection Assays, Nucleic Acid Labeling |
| Product Type | Kit |
| Quantity | 50 Preps |
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Frequently Asked Questions (FAQs)
Poor quality/poor purity RNA is typically due to the following:
- Partially degraded RNA: Use fresh tissue or cells, or use RNAlater Stabilization Solution if tissue cannot be frozen immediately.
- DNA contamination: If the sample contains organic solvents, strong buffers, or an alkaline solution, we recommend performing phase separation at 8 degrees C.
- A low A260/A280 ratio (<1.6): This indicates that an insufficient amount of the TRI Reagent was used. Incubation of the homogenate at room temperature for 5 minutes can help nucleoproteins dissociate from RNA. Phenol contamination could also cause a low absorbance ratio.
The column cut-off size is 200 nt. Using a RiboPure kit is not appropriate for miRNA recovery. For miRNA isolation, we would suggest using a mirVana kit, the TRIzol Plus RNA Purification Kit, or the PureLink RNA Micro Kit.
The PureLink RNA Mini Kit provides rapid column-based purification of total RNA, without organic lysis (phenol/chloroform). You can obtain up to 1,000 µg of purified RNA from a single extraction. The RiboPure RNA Purification Kits combine a phenol/guanidine thiocyanate solution with a glass-fiber filter purification method.
The elution buffer composition is proprietary.
For Research Use Only. Not for use in diagnostic procedures.