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Cat. No.: MFMM-10k (for 10KU) 

Cat. No.: MFMM-1m (for 1MU) 

For M-MLV Reverse Transcriptase active at 65°C, please see Ultra Thermo-stable M-MLV Reverse Transcriptase (RNase H-).

For M-MLV Reverse Transcriptase active at 72°C, please see Prime Thermo-stable M-MLV Reverse Transcriptase (RNase H-).

Description

Mutant M-MLV Reverse Transcriptase (Mutant M-MLV RT) is a variant of M-MLV reverse transcriptase obtained through gene modification and recombinant technology. Compared to conventional M-MLV reverse transcriptase, this enzyme exhibits increased tolerance to inhibitors that interfere with cDNA synthesis, enabling stable and specific cDNA synthesis from samples of various origins. Additionally, it facilitates faster reactions, reducing the incubation time from >50 min to 10 min. Its ability to perform continuous synthesis (with a final product > 12 kb) has also been enhanced compared to previous versions.

Application

This enzyme can be utilized in fluorescence-based RT-RPA or RPA. Depending on primer selection, optimization, and detection methods, the detection limit ranges from 10 to 100 copies per assay, with a reaction time not exceeding 15 minutes.

Quality Control

• Protein Purity: The purity of the reverse transcriptase was assessed using size exclusion high-performance liquid chromatography (SEC-HPLC).
• Endonuclease Activity: Endonuclease activity was assessed by incubating 10 μL of reverse transcriptase with 500 ng of circular plasmid DNA in a 50 μL reaction system at 37°C for 4 hours. The activity was then evaluated using agarose gel electrophoresis.
• Exonuclease Activity: Exonuclease activity was evaluated by incubating 10 μL of reverse transcriptase with 500 ng of double-stranded DNA fragments in a 50 μL reaction system at 37°C for 4 hours. The activity was assessed using agarose gel electrophoresis.
• RNA Hydrolysis Activity: RNA hydrolysis activity was determined by incubating 10 μL of reverse transcriptase with 500 ng of single-stranded RNA in a 50 μL reaction system at 37°C for 4 hours. The activity was evaluated using agarose gel electrophoresis.
• Host DNA Residuals: Residual host DNA content was quantified using fluorescent quantitative PCR with Escherichia coli 16S rDNA as the target sequence.

Storage

Store at -80°C, avoiding repeated freeze-thaw cycles.

Related: 

• Mutant M-MLV Reverse Transcriptase
• Thermo-stable M-MLV Reverse Transcriptase (RNase H-)
• Ultra Thermo-stable M-MLV Reverse Transcriptase (RNase H-)
• Prime Thermo-stable M-MLV Reverse Transcriptase (RNase H-)
• TGI-RTase Reverse Transcriptase
• Warm Activated Reverse Transcriptase

Only for research and not intended for treatment of humans or animals

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