AMV Reverse Transcriptase is both an RNA-dependent and DNA-dependent DNA polymerase purified from Avian Myeloblastosis Virus. It is most commonly used for first- and second-strand cDNA synthesis, but can also be used for dideoxy DNA sequencing, RNA sequencing, primer extension, 3'-end labeling of DNA fragments, generation of single-stranded probes for hybridization, and genomic footprinting. AMV Reverse Transcriptase activity requires either Mg2+ or Mn2+, and a primer of greater than eight nucleotides in length.
Storage Buffer: 200 mM potassium phosphate (pH 7.2), 2 mM DTT, 0.2% Triton-X 100, 50% glycerol.
Storage Conditions: Once thawed, store at -20°C. Aliquots of enzyme may be stored for longer periods at -70°C.
Reaction Buffer (5X): 250 mM Tris-HCl (pH 8.3), 250 mM KCl, 5 mM DTT, 50 mM MgCl2.
Concentration: 20 U/µl
Unit Definition: One unit is defined as that amount of enzyme which incorporates one nanomole of [3H] dTMP into acid-insoluble product in 10 minutes at 37°C with poly (A)+ oligo(dT)12-18 as template/primer.
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