Obtain Higher Yields of cDNA than with the Deletion Mutant

Moloney Murine Leukemia Virus Reverse Transcriptase, RNase H Minus (M-MLV RT [H–]), Point Mutant, is an RNA-dependent DNA polymerase that can be used in cDNA synthesis with long RNA templates (>5kb). The lack of RNase H activity is beneficial for this application, as RNase H can start to degrade templates when incubation times are long, as they may be when synthesizing long cDNAs. Although many researchers are successful in using M-MLV RT (H+) for analytical and some preparative cDNA applications, reverse transcriptases lacking RNase H activity provide another option to prepare long cDNAs and libraries containing a high percentage of full-length cDNA.

Advantages

• RNase H Minus: Provides optimal conditions to prepare full-length cDNA from long RNA templates.
• Temperature Stability: Thermostability of this point mutant minimizes problems associated with secondary structure.
• Increased Polymerase Activity: M-MLV RT (H–), Point Mutant, gives higher yields of cDNA compared with the deletion mutant (Cat.# M5301).
• Provided with 5X Reaction Buffer: 250mM Tris-HCl (pH 8.3 at 25°C), 375mM KCl, 15mM MgCl₂, 50mM DTT.
• Broad Working Range: More tolerance to variations in enzyme and substrate concentrations means improved consistency in performance.

Applications

• First-strand cDNA synthesis.
• Primer extension.