Application

The Titan One Tube RT-PCR Kit is used to analyze RNA (total RNA, poly (A)⁺RNA, viral RNA) from a variety of sources (animal, plant, human, viral, and bacterial). In a single tube, the Titan One Tube RT-PCR Kit will reverse transcribe RNA into cDNA, then amplify the cDNA with a thermal cycling reaction. The enzymes AMV Reverse Transcriptase and the Expand High Fidelity PCR System are used, respectively. Depending on the primers chosen, a specific message or the entire population of transcripts can be amplified. The RT-PCR method also permits the cloning of rare messages without the need to construct cDNA libraries. The kit may be used for:??Qualitative, semi quantitative, or quantitative analysis of RNA transcription levels even in single cells in combination with gel-based detection methods or ELISA-based detection methods
??Cloning of RNA up to 6 kb
??Mutation analysis at RNA level in combination with sequencing or other mutation scanning techniques

Features and Benefits

??Reduces the error rate in PCR due to the proofreading capability. A threefold higher fidelity is obtained in comparison to Taq DNA polymerase.
??Delivers increased sensitivity due to the high efficiency of all three enzymes.
Is faster than conventional two-step RT-PCR.

General description

Download the CofA, SDS and IFU specific to your product lot and format

The Titan One Tube RT-PCR Kit provides reagents, including controls, control primers, nucleotides, RNase inhibitor, and Water, PCR Grade, for performing one-step RT-PCR analysis. In a single tube, the system will reverse transcribe RNA into cDNA, then amplify the cDNA with a thermal cycling reaction. The enzymes AMV Reverse Transcriptase and the Expand High Fidelity PCR System are used, respectively. Depending on the primers chosen, a specific message or the entire population of transcripts can be amplified. The RT-PCR method also permits the cloning of rare messages without the need to construct cDNA libraries.
The Titan One Tube RT-PCR Kit uses Reverse Transcriptase AMV (the enzymatically active forms of the purified enzyme are α, β:β and α:β) for first-strand cDNA synthesis and the Expand High Fidelity enzyme blend, consisting of Taq DNA Polymerase and a polymerase with proofreading activity which leads to increased PCR fidelity. In addition, the kit includes a single optimized RT-PCR buffer, control RNA from a human cell line (K-562) and control primers for human β-actin mRNA.

Other Notes

For life science research only. Not for use in diagnostic procedures.

Packaging

1 kit containing 9 components

Legal Information

Use of this product is covered by US patent claims and corresponding patent claims outside the US. The purchase of this product includes a limited, non-transferable immunity from suit under such patent claims for using only this amount of product for the purchaser′s own internal research. No right under any other patent and no right to perform commercial services of any kind, including without limitation reporting the results of purchaser′s activities for a fee or other commercial consideration, is conveyed expressly, by implication, or by estoppel. This product is for research use only. Diagnostic uses under Roche patent claims require a separate license from Roche. Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.</ br>NOTICE TO PURCHASER: LIMITED LICENSE<br />Use of this product is covered by US Patent No. 6,127,155 and corresponding patent claims outside the US.? The purchase of this product includes a limited, non-transferable immunity from suit under the foregoing patent claims for using only this amount of product for the purchaser′s own internal research.? No right under any other patent claim and no right to perform commercial services of any kind, including without limitation reporting the results of purchaser′s activities for a fee or other commercial consideration, is conveyed expressly, by implication, or by estoppel.? This product is for research use only. Human and veterinary diagnostic uses under Roche patent claims require a separate license from Roche.? All uses other than internal research and human and veterinary diagnostic uses under Roche patent claims require a separate license from Thermo Fisher Scientific. Further information on purchasing licenses from Roche may be obtained by contacting the Licensing Department of Roche Molecular Systems, Inc., 4300 Hacienda Drive, Pleasanton, California 94588, USA or Roche Diagnostics GmbH, Sandhofer Strasse 116, 68305 Mannheim, Germany.? Further information on purchasing licenses from Thermo Fisher Scientific may be obtained by contacting the Licensing Department of? Thermo Fisher Scientific, 5791 Van Allen Way, Carlsbad, California 92008, USA.

Specificity

Specificity of the reaction strongly depends on primer design and DNA contamination.
To exclude artefacts from DNA targets appropriate positive and negative control reaction should be included.
Heat inactivation:?2 min at 94 °C
Reverse Transcriptase AMV