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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.
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 plus||50-200U|
|5×RT Reaction Buffer||5μL|
|dNTP (10mM each)||1.25μL|
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).