Transcriptor High Fidelity cDNA Synthesis Kit

Uses

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 interest
• Sequencing transcriptomes
• Generating cDNA libraries with large and full-length inserts
• Generating gene expression analysis, like RNA splicing analysis

As 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).

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. During this process, reverse transcription produces frequent replication errors. One accepted explanation of this inaccuracy is the lack of RT 3′-5′ exonuclease activity. The naturally high error rate of reverse transcriptases is not optimal for many different applications.