M-MLV Reverse Transcriptase, RNase H Minus

Product Information

Product description: M-MLV Reverse Transcriptase, RNase H Minus is an RNA-dependent DNA polymerase with no detectable RNase H activity. A point mutation in the RNase H domain increases the thermostablity of the enzyme and support greater cDNA yield of full-length transcripts than wild type M-MLV Reverse Transcriptase.

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: 20mM Tris-HCl (pH7.5 at 25℃), 0.1mM EDTA, 200mM NaCl, 0.01% NP-40, 1mM DTT and 50% Glycerol.

Companion Product: 5X RT Reaction Buffer, Cat#ON-067, 250mM Tris-HCl (pH8.3 at 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 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. Shufeng Liu, Stephen P. Goff, et al. FEBS Letters. 580, 1497-1501(2006).

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