RevertAid Premium Reverse Transcriptase

Thermo Scientific RevertAid Premium Reverse Transcriptase was developed through in vitro evolution of M-MuLV RT. The enzyme possesses an RNA-dependent and DNA-dependent polymerase activity, but lacks RNase H activity. The engineered enzyme features dramatically improved thermostability, robustness, and an increased synthesis rate compared to wild type M-MuLV RT.

The eliminated RNase H activity enables the enzyme to produce very long RNA transcripts up to 20 kb. Due to its high thermostability, the enzyme maintains full activity during the entire reverse transcription reaction and generates high yields of cDNA. The reaction temperature can be increased up to 60°C for efficient transcription of RNA regions with a high secondary structure, or to improve specificity using gene-specific primers.

Highlights

• Thermostable – 90% active after incubation at 50°C for 60 minutes in a reaction mixture
• Active up to 60°C
• High yields of full-length cDNA as long as 20 kb
• Efficient – complete cDNA synthesis in 30 minutes
• Incorporates modified nucleotides

Applications

• First strand cDNA synthesis for RT-PCR and RT-qPCR
• Reverse transcription at elevated temperatures to reduce effects of secondary structure
• Synthesis of cDNA for cloning and expression
• Generation of labeled cDNA probes for microarrays
• DNA labeling
• Analysis of RNA by primer extension

Includes

• RevertAid Premium Reverse Transcriptase (200 U/µL)
• 5X RT Buffer