RevertAid H Minus Reverse Transcriptase

Detailní popis

Thermo Scientific RevertAid H Minus Reverse Transcriptase is a  recombinant M-MuLV RT. The enzyme possesses an RNA-dependent and DNA-dependent polymerase activity, but lacks RNase H activity due to a  point mutation in the RNase H domain (see References 1, 2). RevertAid H Minus Reverse Transcriptase does not degrade RNA in RNA-DNA hybrids during synthesis of the first strand cDNA, and therefore high yields of full-length cDNA from long templates are obtained. RevertAid H  Minus Reverse Transcriptase maintains activity over a wide temperature range (42 to 50°C).

Highlights

• High yields of full-length first strand cDNA up to 13 kb
• Optimum activity at 42 to 45°C
• Active up to 55°C
• Incorporates modified nucleotides (e.g., Cy3-, Cy5-, rhodamine-, aminoallyl-, fluorescein-labeled nucleotides)

Applications

• First strand cDNA synthesis for RT-PCR and real-time RT-PCR (see Reference 3)
• 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 (see​ Reference 3)
• Analysis of RNA by primer extension (see​ Reference 3)

Includes

• RevertAid H Minus Reverse Transcriptase (200 U/μL) is supplied in storage buffer including: 50 mM Tris-HCl (pH 7.5), 0.1 M NaCl, 1 mM EDTA, 5 mM DTT, 0.1% (v/v) Triton X-100, and 50% (v/v) glycerol
• 5X Reaction Buffer: 250 mM Tris-HCl (pH 8.3 at 25°C), 250 mM KCl, 20 mM MgCl₂, 50 mM DTT)

Note