Promega, M3683, M-MLV Reverse Transcriptase, RNase H Minus, Point Mutant

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 MgCl 2 , 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.
Thermal stability and synthesis of large transcripts. cDNAs were synthesized from 1µg of total human RNA using M-MLV (H–), Point Mutant ( Panel A ), or SuperScript® II ( Panel B ) reverse transcriptase at 37, 42 and 50°C. GoTaq® Long PCR amplifications were performed on each transcription reaction using five primer sets, amplifying products from 9.4kb to 1.5kb. Ten microliters of the 9.4, 6.9 and 5.2kb PCR products and 1µl of the 3.2 and 1.5kb PCR products were separated on a 1% agarose gel. Lane M. BenchTop 1kb DNA Ladder (Cat.# G7541).