Avian myeloblastosis virus reverse transcriptase (or AMV reverse transcriptase) catalyses the polymerization of DNA using DNA, RNA or RNA: DNA hybrids. It requires a primer (DNA primers are more effective than RNA primers) as well as Mg2 + or Mn2 + ions. The enzyme has an intrinsic activity of RNase H. Nonionic detergents and sulfhydryl compounds stabilize its enzymatic activity in vitro.

The mature αβ form, the most active form of the AMV reverse transcriptase, has multiple enzymatic activities including RNA-dependent DNA polymerase, DNA-dependent DNA polymerase, RNase H and unwinding activity. AMV reverse transcriptase is used for cDNA synthesis, for first strand cDNA synthesis for use in subsequent amplification reactions and for dideoxy DNA sequencing.

The enzyme can also be used for RNA sequencing, 3 'labeling of DNA fragments and generation of ss probes for genomic fingerprints.