RNA Extraction for Real-Time PCR
Optimal RNA extraction is a key to success for any RT-qPCR experiment. To avoid false positives and background, the complete removal of genomic DNA is another key variable. Ambion® products will simplify your experiments. You can even get RT-qPCR results direct from cells without prior RNA isolation with Ambion® Cells-to-CT™ kits
Which RNA Isolation Kit is Right for Your qRT-PCR Experiment?
| Gold standard for highly pure, intact RNA | High-quality RNA in less than 20 minutes | High-throughput purification of RNA and DNA | Complete, no purification system for qRT-PCR results | |
|---|---|---|---|---|
| Order now | Order now | Order now | Order now | |
| Trizol® Reagents | PureLink™ Kits | MagMAX™ Kits | Cells to Ct™ Kits | |
| Recommendations | Most Cited, Top Seller | Best for qRT-PCR | ||
| Isolation method | Organic reagent | Silica filter, column format (convenience) | Magnetic beads (scalability, flexibility) | Chemical lysis (time to qRT-PCR results) |
| High throughput compatible | No | Yes (plate format available) | Yes | Yes |
| RT reagents included | No | No | No | Yes |
| qPCR reagents included | No | No | No | Yes |
| Prep time | ~1hr | < 20 minutes | ~45 minutes | 10 minutes |
| Compatible sample types | Most sample types, particularly more difficult to lyse | Cells and tissues | Cells, blood, plants | Cells |
| Amount of starting material | 100 mg of tissue or 107 cells (requires 1ml reagent) | Up to 200 mg tissue, 5x106 to 1 x 108 cells | Up to 100 mg tissue, Up to 5x106 cells | 1–10,000 cells |
| Price per prep range | $1.40 - $7.42 | $4.50 - $6.03 | $3.00 - $8.00 | $9.62 - $19.00 |
Eliminate Genomic DNA Contamination
Genomic DNA contamination can lead to false positive RT-PCR results. We offer a variety of Ambion® tools for minimizing genomic DNA contamination from RNA samples prior to RT-PCR. For example, DNA-free™ DNase Treatment and Removal Reagents are designed for removing contaminating DNA from RNA samples and for the removal of DNase after treatment without Proteinase K treatment and organic extraction. In addition, we offer TURBO DNase™ enzyme kits, a hyperactive enzyme engineered from wild-type bovine DNase. The proficiency of TURBO DNase™ enzyme in binding very low concentrations of DNA means that the enzyme is particularly effective in removing trace quantities of DNA contamination. To prevent cross contamination during PCR experiments, we also offer DNAZap™ DNA Degradation Solution and RNase-free barrier pipette tips.
Technical Resources
- Advanced Real-Time PCR Technology - TechNotes 14(4)
- Cells-to-cDNA™ II Using FACS Sorted Cells - TechNotes 9(3)
- Cells-to-cDNA™II Applications 9(4) - TechNotes 9(4)
- Getting Started with microRNA Expression Analysis Research using Real-Time PCR - TechNotes 13(4)
- Go from Cells in Culture to Real-Time PCR Data Even Faster - TechNotes 15(2)
- Go From Cells to cDNA Directly: Using FACS Samples - TechNotes 9(3)
- Matched siRNAs and Assays: Ambion® + Applied Biosystems® = RNAi Success - TechNotes 11(4)
- miRNA Profiling: Save Time and Sample Using Real-Time PCR - TechNotes 14(2)
- Quantitating Gene Expression Directly from Cell Lysates Using TaqMan® Real-Time PCR Analysis - TechNotes 14(1)
- Quick Real-time Analysis of Small Cell Samples - TechNotes 9(2)
- Rapidly Optimize Anti-miR™ Inhibitor Transfection Using Real-Time RT-PCR - TechNotes 14(4)
- Ready for Real-time in 5 Minutes - TechNotes 11(4)
- Real-time PCR Goes Prime Time - TechNotes 8(1)
- Real-Time Relative RT-PCR: How It's Done - TechNotes 8(5)
- Real-Time RT-PCR Directly from Cell Lysates: A Complete Workflow without RNA Purification - TechNotes 14(3)
- Streamline the Path from RNA to Real-Time PCR Results - TechNotes 15(3)
- The Best Online Resource for siRNAs and Real-Time PCR Assays Just Got Better - TechNotes 15(3)
- Using SYBR® Green for Real-Time RT-PCR? Try SYBR Green Cells-to-CT™ Kits - TechNotes 15(3)