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Invitrogen™ RiboPure™ RNA Purification Kit, blood
Description
Includes
25mL Elution Solution; 40mL Lysis Solution; 25mL Acid-Phenol:Chloroform; 5mL Sodium Acetate Solution; 35mL Wash Solution 1; 70mL Wash 2/3 Concentrate; 100mL RNAlater Solution; 40 Filter Cartridges; 80 Collecion Tubes; 100μL DNase I, 8 U/μL; 850μL 20X DNase Buffer; 1.2mL DNase Inactivation Reagent; 1mL Formaldehyde Load Dye. This kit contains enough reagents for 40 RNA isolation from 300–500μL whole blood.
The RiboPure™-Blood Kit is for the isolation of high-quality RNA directly from whole blood. The kit combines two methods of RNA purification to efficiently remove blood components (protein, heme, genomic DNA, and RNases) that can compromise downstream procedures. The kit includes sufficient reagents for 40 RNA purifications from whole blood samples ranging from 0.3–0.5 mL each.
Advantages of the RiboPure™-Blood Kit:
• Purify total RNA directly from whole blood—without prior recovery of WBCs
• Simple, fast procedure—from blood to RNA in less than 30 minutes
• Includes RNAlater™ Solution—store blood samples 3 days or more at room temperature
• Includes DNA-free™ Reagents—to ready RNA for RT-PCR and array analysis
Applications
The RiboPure™-Blood Kit produces high yields of pure RNA directly from whole blood, eliminating the need to isolate WBCs prior to RNA purification. RNA recovered using the RiboPure™-Blood Kit can be used in the most challenging applications, including real-time RT-PCR and array analysis. Average yields of RNA are between 2–4 μg/0.5 mL of whole blood (variation is due to differences in WBC numbers between samples).
Stabilize RNA in whole blood
When collecting blood samples for RNA analysis, storage of the samples prior to RNA isolation is often required. The RiboPure™-Blood Kit contains RNAlater™ Solution for the stabilization of RNA in whole blood. It also “freezes” the gene expression profile of the cells. Treated samples can be safely stored at ambient temperature for extended periods of time (up to three days or more). Blood samples stored in RNAlater™ Solution yield RNA of comparable quality to blood samples processed directly (see figure).
Simple procedure yields highly pure RNA
The RiboPure™-Blood procedure consists of three steps: 1) addition of Lysis Solution directly to RNAlater™-treated or fresh whole blood, 2) initial RNA purification by phenol/chloroform extraction, and 3) final RNA purification on a glass fiber filter. An additional DNase treatment can then be performed with the DNA-free™ Reagents included in the kit. The entire procedure, including DNase treatment, can be completed in less than 1 hour.
Accessory products
The RNA can subsequently be enriched for white blood cell (WBC) mRNA using our GLOBINclear™ Kits to selectively remove globin transcripts. The Mouse RiboPure™-Blood RNA Isolation Kit (Cat. No. AM0951) is also available for the recovery of total RNA—and optionally microRNA—directly from mouse or rat blood. Mouse RiboPure™-Blood includes sample tubes preloaded with RNAlater™ Solution to prevent RNA degradation and preserve RNA profiles.
Specifications
Specifications
| Content And Storage | • 25 mL Elution Solution (4°C) • 40 mL Lysis Solution (4°C) • 25 mL Acid-Phenol:Chloroform (4°C) • 5 mL Sodium Acetate Solution (4°C) • 35 mL Wash Solution 1 (room temperature/4°C) • 70 mL Wash 2/3 Concentrate (room temperature/4°C) • 100 mL RNAlater™ Solution (room temperature) • 40 Filter Cartridges (room temperature) • 80 Collecion Tubes (room temperature) • 100 μL DNase I, 8 U/μL (-20°C, frost-free) • 850 μL 20X DNase Buffer (-20°C) • 1.2 mL DNase Inactivation Reagent (-20°C) • 1 mL Formaldehyde Load Dye (-20°C) This kit contains enough reagents for 40 RNA isolation from 300-500 μL whole blood. |
| Format | Filter Cartridge, Tube |
| Isolation Technology | Organic Extraction, Spin Column |
| Purification Time | 30 min. |
| Sample Type | Blood |
| Final Product Type | Total RNA |
| High-throughput Compatibility | Not High-throughput Compatible (Manual) |
| For Use With (Application) | Microarray Analysis, Real-Time Quantitative PCR (qPCR), Reverse Transcriptase PCR (RT-PCR), Northern Blotting |
| No. of Reactions | 40 Preps |
| Product Line | Ambion, RiboPure |
| Show More |
Frequently Asked Questions (FAQs)
The former (Cat. No. AM1928) is for isolation of RNA from human blood and the latter (Cat. No. AM1951) is for isolation of RNA from mouse and rat blood. Both kits have the same columns but differ in protocol and the wash 1 buffer, wash 2/3 buffer, and sodium acetate solutions. We do not recommend using the mouse kit for human blood samples. The mouse kit accommodates the difference in viscosity of mouse/rat vs. human blood and also the higher mouse and rat blood RNA yield. The mouse kit allows recovery of either total RNA containing small RNAs or total RNA depleted of small RNAs based on which protocol in the manual was followed. With the RiboPure Blood Purification Kit, using the standard protocol in the manual, you can isolate total RNA depleted of small RNAs, but with an alternate protocol, you can isolate total RNA containing small RNAs. Alternatively, if you need small RNAs alone, then you should follow the above alternate protocol for isolation of total RNAs containing small RNAs, and then follow Section IVA in the mirVana miRNA Isolation Kit manual (Cat. No AM1560).
Its composition is 0.1 mM EDTA in nuclease-free water.
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.
Please visit our website for tips for working with blood samples.
For Research Use Only. Not for use in diagnostic procedures.
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