Titan One Tube RT-PCR Kit
The Titan One Tube RT-PCR Kit is designed for fast, sensitive and reproducible analysis of RNA by high fidelity reverse transcription polymerase chain reaction (RT-PCR) in a one-step reaction.
Titan™ One Tube RT-PCR Kit uses Avian Myeloblastosis Virus (AMV) reverse transcriptase for first strand cDNA synthesis and the Expand™ High Fidelity enzyme blend consisting of Taq DNA polymerase and Tgo DNA polymerase for amplification of cDNA by PCR. Tgo DNA polymerase possesses proofreading activity leading to an increased PCR fidelity. This kit reduces the error rate, allows the amplification of fragments up to 6 kb by RT-PCR, delivers increased sensitivity. It is considered faster than conventional two-step RT-PCR.
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
Titan One Tube RT-PCR Kit has been used:
• to complete cDNA synthesis in reverse transcription-quantitativ
• to clone sirtuin 6 (SIRT6) into the backbone
of pDsRed2-N1[3]
• to amplify full-length and truncated cDNA fragments of C-X-C motif chemokine 5 (CXCL5)[4]
• 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
• for cloning of RNA up to 6 kb
• for mutation analysis at RNA level in combination with sequencing or other mutation scanning techniques
• 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.
1 kit containing 9 components
