Reverse Transcriptase with Stability at Higher Temperatures

Avian Myeloblastosis Virus Reverse Transcriptase (AMV RT) catalyzes DNA polymerization using template DNA, RNA or RNA:DNA hybrids. It requires a primer (DNA primers are more efficient than RNA primers) as well as Mg²⁺ or Mn²⁺. The enzyme possesses an intrinsic RNase H activity. Both nonionic detergents and sulfhydryl compounds stabilize the enzyme activity in vitro.

Features

• High Concentration: Cat.# M9004 contains 600 units of AMV RT at 20–25u/μl.
• 5X Reaction Buffer: 250mM Tris-HCl (pH 8.3 at 25°C), 250mM KCl, 50mM MgCl₂, 2.5mM spermidine, 50mM DTT.
• Temperature Stability: AMV RT is the preferred reverse transcriptase for templates with high secondary structure due to its stability at higher reaction temperatures (37–58°C).

Applications

• First- and second-strand synthesis of cDNA.
• Primer extensions and RNA sequencing.
• RT-PCR. Up to 10μl of an RT reaction containing AMV RT and the supplied AMV RT Reaction Buffer can be added to a 50μl PCR amplification reaction that uses Taq DNA polymerase. If GoTaq® DNA Polymerase or PCR Master Mix are used, up to 25μl of an RT reaction can be added per 50μl PCR.

Further Reading

1. Kacian, D.L. (1977) Meth. Virol. 6, 143.
2. Mierendorf, R.C. and Pfeffer, D. (1987) Meth. Enzymol. 152, 563–6.