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Real-Time PCR RNA Quantification Workflow
RNA quantification using reverse transcription, real-time PCR (RT-qPCR) is a powerful, sensitive, and precise method. RT-qPCR is capable of high throughput, and is relatively easy to perform.
RNA isolation is followed by cDNA synthesis – if you include a prior RNA quantitation step, you’ll be better able to optimize your qPCR analysis.
RT-qPCR is a multi-step protocol that requires both high quality RNA as input material, and optimal conversion of this RNA to a DNA copy for accurate real-time PCR detection (qPCR).
Make sure you optimize each step. Click the tabs above to find out more and see the available products.
The sensitivity and accuracy of RT-qPCR detection will depend on the quality and quantity of your input cDNA – which in turn depends on the quality and integrity of the isolated RNA. To avoid false positives and background, the removal of genomic DNA is another key variable.
Ambion® RNA purification products will simplify your RT-qPCR experiments. Find out more about RNA Extraction for Real-Time PCR. You can also get RT-qPCR results direct from cells with Ambion® Cells-to-CT™.
Fresh/frozen cells or tissue samples
| Rapid & relaible method | High-throughput purification | microRNA from most samples | RNA from precious micro-sized samples | |
|---|---|---|---|---|
| Order Now | Order Now | Order Now | Order Now | |
| PureLink™ RNA Mini Kit | MagMAX™-96 Total RNA Isolation Kit | mirVana™ miRNA Isolation Kit | RNAqueous®-Micro Kit | |
| RNA type(s) isolated | large RNA molecules only (mRNA and rRNA) | large RNA molecules only (mRNA and rRNA) | Small & large RNA molecules (microRNA, tRNA, mRNA, rRNA) | Small & large RNA molecules (microRNA, tRNA, mRNA, rRNA) |
| Isolation method | Fast, convenient silica column | Scalable, flexible format with magnetic beads | Highest purity and convenience; includes both organic extraction and silica column | Fast, convenient low elution silica column |
| Prep. time | 20 minutes | <45 minutes | 30 minutes | 15 minutes |
| Amount of starting material | 10 mg to 100 mg of tissue Up to 5 x 107 cells | Up to 10 mg of tissue Up to 100,000 cells | Up to 100 mg of tissue Up to 1 x 107 cells | Up to 10 mg of tissue Up to 100,000 cells |
The Qubit® and Quant-iT™ RNA assays provide specific and sensitive RNA or microRNA quantitation - even at low concentrations – with no interference from contaminants. Unlike UV absorbance readings which are indiscriminate, these assays can distinguish between DNA, RNA, proteins, and free nucleotides, Higher sensitivity than UV absorbance readings makes it possible to measure low-abundance samples. Intuitive format makes it easier and faster to get your quantitation results. Find out more about Qubit® and Quant-iT™ technology for your precious samples. |  |
1-20 samples: Qubit® Assays for use on Qubit® Fluorometer
20-2000 samples: Quant-iT™ Assays for use on fluorescent microplate reader
For reliable generation of first strand cDNA, SuperScript® VILO™ cDNA synthesis kit brings you proven SuperScript® III Reverse Transcriptase formulated in an enhanced buffer system. Reduce amplification bias from low- or high- abundance gene products, and get data that exhibit superior linearity across broad range on input material. The SuperScript® VILO™ cDNA kit offers consistent result for your RT experiments and is also available in a convenient single-tube format – allowing simplifed, time-saving workflow and reduced risk of contamination. |  |
RT-qPCR is a multi-step protocol that requires both high quality RNA as input material, and optimal conversion of this RNA to a DNA copy for accurate real-time PCR detection (qPCR).
Make sure you optimize each step. Click the tabs above to find out more and see the available products.
The sensitivity and accuracy of RT-qPCR detection will depend on the quality and quantity of your input cDNA – which in turn depends on the quality and integrity of the isolated RNA. To avoid false positives and background, the removal of genomic DNA is another key variable.
Ambion® RNA purification products will simplify your RT-qPCR experiments. Find out more about RNA Extraction for Real-Time PCR. You can also get RT-qPCR results direct from cells with Ambion® Cells-to-CT™.
Fresh/frozen cells or tissue samples
| Rapid & relaible method | High-throughput purification | microRNA from most samples | RNA from precious micro-sized samples | |
|---|---|---|---|---|
| Order Now | Order Now | Order Now | Order Now | |
| PureLink™ RNA Mini Kit | MagMAX™-96 Total RNA Isolation Kit | mirVana™ miRNA Isolation Kit | RNAqueous®-Micro Kit | |
| RNA type(s) isolated | large RNA molecules only (mRNA and rRNA) | large RNA molecules only (mRNA and rRNA) | Small & large RNA molecules (microRNA, tRNA, mRNA, rRNA) | Small & large RNA molecules (microRNA, tRNA, mRNA, rRNA) |
| Isolation method | Fast, convenient silica column | Scalable, flexible format with magnetic beads | Highest purity and convenience; includes both organic extraction and silica column | Fast, convenient low elution silica column |
| Prep. time | 20 minutes | <45 minutes | 30 minutes | 15 minutes |
| Amount of starting material | 10 mg to 100 mg of tissue Up to 5 x 107 cells | Up to 10 mg of tissue Up to 100,000 cells | Up to 100 mg of tissue Up to 1 x 107 cells | Up to 10 mg of tissue Up to 100,000 cells |
The Qubit® and Quant-iT™ RNA assays provide specific and sensitive RNA or microRNA quantitation - even at low concentrations – with no interference from contaminants. Unlike UV absorbance readings which are indiscriminate, these assays can distinguish between DNA, RNA, proteins, and free nucleotides, Higher sensitivity than UV absorbance readings makes it possible to measure low-abundance samples. Intuitive format makes it easier and faster to get your quantitation results. Find out more about Qubit® and Quant-iT™ technology for your precious samples. | |
1-20 samples: Qubit® Assays for use on Qubit® Fluorometer
20-2000 samples: Quant-iT™ Assays for use on fluorescent microplate reader
For reliable generation of first strand cDNA, SuperScript® VILO™ cDNA synthesis kit brings you proven SuperScript® III Reverse Transcriptase formulated in an enhanced buffer system. Reduce amplification bias from low- or high- abundance gene products, and get data that exhibit superior linearity across broad range on input material. The SuperScript® VILO™ cDNA kit offers consistent result for your RT experiments and is also available in a convenient single-tube format – allowing simplifed, time-saving workflow and reduced risk of contamination. | |