Thermo Scientific Maxima Reverse Transcriptase (RT) through in vitro evolution of M-MuLV RT have to. The enzyme has both RNA-dependent and DNA-dependent polymerase activity but lacks RNase H activity due to mutations in the RNase H domain of M-MuLV RT. Compared with wild-type M-MuLV RT, the engineered enzyme has significantly improved thermal stability, 50-fold higher processivity, robustness, and improved synthesis rate.

Elimination of RNase H activity enables the enzyme to produce RNA transcripts up to 20 kb long. Due to its high thermal stability, the enzyme maintains full activity throughout the reverse transcription reaction and produces high yields of cDNA. The reaction temperature can be increased to 65°C for efficient transcription of RNA with complex secondary structures, or gene-specific primers can be used to increase specificity. Maxima H Minus enzymes are extremely productive and provide increased resistance to common reaction inhibitors such as guanidine, formamide and ethanol.

Product Advantages

• Thermal stability—90% activity after 60 minutes of incubation in the reaction mixture at 50°C.
• Maximum activity temperature 65°C
• High yield of full-length cDNA, up to 20 kb
• High sensitivity—can be performed from a wide range of total RNA amounts (1 pg to 5 μg) Reproducible cDNA synthesis
•Efficient—cDNA synthesis is completed in 15 to 30 minutes
•Enhanced resistance to common reaction inhibitors
•Incorporated modified nucleotides

Applications

•First-strand cDNA synthesis for RT-PCR and real-time RT-PCR.
•Perform reverse transcription at high temperatures to reduce the impact of secondary structure.
•Synthesis of cDNA for cloning and expression.
•Generation of labeled cDNA probes for use in microarrays.
•Analysis of RNA by primer extension.

Note
•Also available::Maxima H Minus First Strand cDNA Synthesis Kit.