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M-MLV Reverse Transcriptase, RNase H Plus

  • Cat.No.ON-076
  • SourceRecombinant E.coli
  • Purity>95% by SDS-PAGE
  • Storage-20℃
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Product description: M-MLV Reverse Transcriptase,RNase H Plus is an RNA-dependent DNA polymerase that can be used in cDNA synthesis with long messenger RNA templates. M-MLV Reverse Transcriptase,RNaseH Plus will also extend primers hybridized to single-stranded DNA. Second strand cDNA synthesis can be achieved from some mRNA templates without an additional DNA polymerase.

Source: Recombinant E.coli

Concentration and Size: 200U/μL

Unit Definition: One unit is defined as the amount of enzyme required to catalyze the transfer of 1nmol of deoxynucleotides into acid-precipitable material in 10 minutes at 37℃.

Storage Buffer: 50mM Tris-HCl (pH7.5, 25℃), 0.1mM EDTA, 100mM NaCl, 0.1%Triton X-100, 1mM DTT and 50% Glycerol.

Companion Product: 5×RT Reaction Buffer. Cat#ON-067, 250mM Tris-HCl (pH8.3, 25℃),375mM KCl, 15mM MgCl2, 50mM DTT.

Related Products:

M-MLV Reverse Transcriptase, RNase H Minus, Cat#ON-047

Ribonuclease Inhibitor, Human Placenta, Cat#ON-039

5×RT Reaction Buffer, Cat#ON-067

Before the first-strand Synthesis of DNA, heat the primer(about 0.5μg) and  template(up to 2μg) to 70℃ for 5min to melt secondary structure within the template. Cool the tube on ice to prevent secondary structure from reforming.

2. Add the following components to the annealed primer/template:

   

M-MLV reverse transcriptase, RNase H plus50-200U
5×RT Reaction Buffer5μL
RNasin20U
dNTP (10mM each)1.25μL
Total25μL

3. Incubate for 60min at 37℃ for random primers or 42℃ for other primers. The extension temperature may be optimized between 37℃ and 42℃.

Storage Conditions: -20℃

Quality Assurance: Free of endonuclease, exonuclease, Nickase and RNase activities.

Physical Purity: >95% by SDS-PAGE.

1. Yu Chen, Weiguo Xu,Qining Sun, Biotechnol Lett. 31, 1051–1057 (2009).

2. Monica J. Roth, Naoko Tanese, and Stephen P. Goff, The Journal Of Biological Chemistry. 260, 9326-9335(1985).

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