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Transcriptor Reverse Transcriptase

Increase Power and Sensitivity in cDNA Synthesis with the new Transcriptor Reverse Transcriptase

With the new recombinant Transcriptor Reverse Transcriptase, you will
  • Achieve high sensitivity in one- or two-step RT-PCR
  • Obtain full-length cDNA transcripts
  • Reverse transcribe difficult templates with high secondary structure
  • Efficiently label RNA
Transcriptor Reverse Transcriptase is the product of choice to transcribe RNA (mRNA, total RNA, viral RNA, and in vitro transcribed RNA) from a variety of sources for the following applications:
  • Synthesis of first-strand cDNA for use in subsequent amplification reactions, using conventional thermal cyclers and real-time PCR instruments (e.g., the LightCycler Instrument)
  • RT-PCR of GC-rich RNA templates with high secondary structure
  • Incorporation of Cy3-, Cy5-, DIG-, Biotin-, and aminoallyl-modified nucleotides during cDNA synthesis (e.g., for use in microarray applications)
  • Retrieval and cloning of the 5- and 3-termini of mRNA by RACE
  • Generation of cDNA libraries with large inserts
Transcriptor Reverse Transcriptase will allow you to Get the most out of your precious RNA samples
  • Improve the overall performance of existing assays.
  • Perform assays that were not previously possible.
Enjoy working with RNA

  • Discover the most powerful combination available today.
Combine: Protector RNase Inhibitor, Transcriptor Reverse Transcriptase, the FastStart PCR Product Family, and PCR-Grade Nucleotides to guarantee your RNA molecules are optimally protected, transcribed, and amplified.

  • Avoid repeating the same experiments and performing multiple RNA extractions.
  • Celebrate quick results.
Transcriptor Reverse Transcriptase is a new recombinant reverse transcriptase expressed in E. coli. The enzyme has RNA-directed DNA polymerase activity, DNA-dependent DNA polymerase activity, unwinding activity and, very importantly, RNase H activity that degrades RNA in RNA:DNA hybrids. If the RNA template is not degraded after first-strand cDNA synthesis, it can bind to the newly-synthesized cDNA and restrict its accessibility to primers during subsequent PCR amplification. RNase H mediated template destruction can prevent this problem and improve the sensitivity of RT-PCR analysis

Transcriptor Reverse Transcriptase optimally combines the individual enzymatic activities, resulting in a number of unique and beneficial properties:

Increase sensitivity in one- or two-step RT-PCR on conventional thermal cyclers
  • Permits the analysis of low-expressed genes and RNA, when limited amounts of sample material are available, using two-step RT-PCR:
In combination with the new FastStart High Fidelity PCR System, as little as 100 pg of total RNA is detected (Figure 1).

Result: The mouse -actin transcript is detected in as little as 100 pg mouse liver total RNA, by two-step RT-PCR.

  • Permits the analysis of low-expressed genes and RNA, when limited amounts of sample material are available, using two-step RT-PCR:
In combination with the Expand High Fidelity PCR System, your target can be detected in as little as 1 pg total RNA and 100 fg mRNA.

Increase sensitivity in two-step quantitative RT-PCR

Analyze genes with very low or extremely high expression levels
  • Transcriptor Reverse Transcriptase has a broad dynamic detection range of 12 to 1.2 x 109 copies of in vitro transcribed RNA, allowing linear and accurate quantification over 108-fold range of input RNA (Figure 2).

Result: Transcriptor Reverse Transcriptase enables a broad dynamic detection range of 12 to 1.2 x 109 copies PBGD per capillary, which equates to a linear quantification of over 108-fold of input RNA.

Generate clear and interpretable results
  • Compared to other products, Transcriptor Reverse Transcriptase delivers high fluorescence intensities and well-shaped curves with identical distances for RNA dilutions (Figure 3).

Result: Transcriptor Reverse Transcriptase produces curves with better shape, higher fluorescence intensity, and lower crossing points compared to other reverse transcriptases

Obtain full-length transcripts during cDNA synthesis

  • Reverse transcription of mRNA up to 14 kb in length is possible, making it the product of choice for very long eukaryotic mRNA (Figure 4).
  • To achieve high sensitivity and yield in the subsequent PCR, use FastStart Taq DNA Polymerase or the new FastStart High Fidelity Enzyme Blend (both products allow hot-start PCR). The Expand Long Template PCR System is recommended for the amplification of long fragments (> 5 kb).

Result: Transcriptor provides high sensitivity and yield when reverse transcribing very long messages

Enjoy the power to reverse transcribe the most difficult templates
  • Due to the very high thermostability of Transcriptor Reverse Transcriptase (up to 65C), robust reverse transcription of GC-rich RNA with high secondary structure can now be easily achieved (Figure 5).
  • In combination with FastStart Taq DNA Polymerase and the new FastStart High Fidelity Enzyme Blend, Transcriptor Reverse Transcriptase is the product of choice to reverse transcribe and amplify difficult RNAs

Result: Transcriptor Reverse Transcriptase has very high thermostability and can be used at temperatures up to 65C

Shorten the reaction time
  • No need to perform an additional RNase H incubation step after reverse transcription (as is recommended for maximum sensitivity when using a RNase H--mutated reverse transcr iptase). This time-consuming, expensive step can be eliminated with Transcriptor Reverse Transcriptase.
  • No need to perform an additional RNase H incubation step after reverse transcription (as is recommended for maximum sensitivity when using a RNase H--mutated reverse transcriptase). This time-consuming, expensive step can be eliminated with Transcriptor Reverse Transcriptase.

Efficiently label RNA
  • Transcriptor Reverse Transcriptase efficiently incorporates modified nucleotides such as Cy3-, Cy5-, aminoallyl-, DIG-, and Biotin-dUTP modified nucleotides, making it an important enzyme in, for example, microarray applications.

Result: Transcriptor Reverse Transcriptase exhibits excellent efficiency for cDNA labeling with Cy3 and Cy5.

Rely on a multi-purpose enzyme, tested in a wide variety of assays
  • Stringent quality criteria are used during the production and manufacturing of Transcriptor Reverse Transcriptase. Each lot is function-tested with conventional thermal cyclers and the LightCycler Instrument, ensuring functionality in end-point and real-time PCR.
  • Additional specifications (e.g., testing for bioburden) increase the safety level when working with RNA.



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