Application
The Transcriptor High Fidelity cDNA Synthesis Kit is designed to reverse transcribe RNA with increased fidelity compared to other reverse transcriptases. The kit features Transcriptor High Fidelity Reverse Transcriptase, a blend of a recombinant reverse transcriptase and a proofreading mediating enzyme optimized for two-step RT-PCR, making this the product of choice for:Cloning genes of interestSequencing transcriptomesGenerating cDNA libraries with large and full-length insertsGenerating gene expression analysis, like RNA splicing analysisAs the kit is also tested with the LightCycler® Instruments and other real-time PCR instruments, the product is also ideal for quantitative RT-PCR applications that require high fidelity.The kit contains all components required for synthesizing cDNA suitable for direct use in qualitative RT-PCR with conventional thermal cyclers or quantitative RT-PCR on real-time PCR instruments. The 50-reaction pack size also includes 10 control reactions (control RNA and control primer mix).
Components
Transcriptor High Fidelity cDNA Synthesis Kit includes the following: Transcriptor High Fidelity Reverse Transcriptase (blend of a recombinant reverse transcriptase and a proofreading mediating enzyme) Transcriptor High Fidelity Reaction Buffer, 5x concentrated Protector RNase Inhibitor dNTP mix, PCR Grade Anchored-oligo (dT)18Primer Random Hexamer Primer DTT Water, PCR Grade Control RNA – total RNA fraction purified from the immortalized cell line K-562* Control PBGD primer mix (forward and reverse primer)** Included only in the 50-reaction pack size.
Features and Benefits
Increase accuracy during reverse transcription reactions. Benefit from an optimized enzyme blend with 7-fold higher fidelity compared to other commonly used reverse transcriptases. Rely on true-fidelity data, as determined by the sequencing of several million bases with the Genome Sequencer 20 System.Combine accuracy with high sensitivity, high yield, and full-length transcripts. Generate full-length transcripts up to 14 kb with the anchored-oligo (dT)18 primers included in the kit . Obtain excellent yields. Detect low-copy number templates; reverse transcribe from as little as 10 pg template RNA. Transcribe a variety of templates, even the most difficult ones (e.g., GC-rich RNAs with high secondary structure) at temperatures up to +55°C.Get results faster. Reduce reverse transcription reaction time to as little as 10 minutes. Be first to finish
General description
Retroviral reverse transcriptases commonly used for cDNA synthesis exhibit a higher error rate than other DNA polymerases used in nucleic acid analysis techniques. This lack in accuracy leads to a significant number of base exchanges or frameshifts, which are further propagated in subsequent PCR reactions. High fidelity (proofreading) PCR enzymes have been available for many years; Roche’;s new high accuracy reverse transcriptase is further able to synthesize high yields of full-length cDNA.High mutation rate is a hallmark of retrovirus replication. This originates in the mechanism of genome replication by the viral-encoded reverse transcriptase, which converts the genomic RNA of the virus to a dsDNA