M-MLV Reverse Transcriptase, RNase H Plus

Product Information

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

Key points of operation

1. 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:

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

Security Information

Storage Conditions: -20℃

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

Physical Purity: ＞95% by SDS-PAGE.

Reference

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).