Skip to content

M-MLV Reverse Transcriptase, RNase H Minus

by Promega
Login to view price.
Size
  • Overview
  • Protocols
  • Specifications
  • Resources
  • First-Strand cDNA Synthesis with Long mRNA Templates

    Moloney Murine Leukemia Virus Reverse Transcriptase, RNase H Minus (M-MLV RT [H–]), is an RNA-dependent DNA polymerase that can be used in cDNA synthesis with long messenger RNA templates (>5kb). This form of M-MLV Reverse Transcriptase has been genetically altered to remove the associated RNase H activity. 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.
    • Provided with 5X Reaction Buffer: 250mM Tris-HCl (pH 8.3 at 25°C), 375mM KCl, 15mM MgCl2, 50mM DTT.
    • May Be Heat-Inactivated: M-MLV RT is inactivated by heating at 70°C for 10 minutes.

    Applications

    • First-strand cDNA synthesis.
    • Primer extension.

    Technical Details

    Storage Buffer: 20mM Tris-HCl (pH 7.5 at 25°C), 0.2M NaCl, 0.1mM EDTA, 1mM DTT, 0.01% Nonidet® P-40 and 50% glycerol.

    Source: Recombinant E. coli strain.

    QC Tests: Activity, SDS-PAGE/purity, DNase, RNase, endonuclease, first-strand cDNA synthesis.

    Unit Definition: One unit is the amount of enzyme required to incorporate 1nmol of deoxynucleotide into acid-precipitable material in 10 minutes at 37°C. The reaction conditions are: 50mM Tris-HCl (pH 8.3 at 25°C), 7mM MgCl2, 40mM KCl, 10mM DTT, 0.1mg/ml BSA, 0.5mM [3H]dTTP, 0.025mM oligo(dT), 0.25mM poly(A) and 0.01% NP-40.

  • Protocols

    Complete Protocol

    Download PDF

    M-MLV Reverse Transcriptase, RNase H Minus Protocol

    PDF (109 KB)

  • Certificate of Analysis

    Lookup Certificate of Analysis

    Storage Conditions

    -30C TO -10C

    Use Restrictions

    For Research Use Only. Not for Use in Diagnostic Procedures.

  • Resources

    Articles

    Citations