AMV Reverse Transcriptase

Avian Myeloblastosis Virus (AMV) Reverse Transcriptase is an RNA directed DNA polymerase which can synthesize a complementary DNA strand initiating from a primer using either RNA (cDNA synthesis) or single-stranded DNA as a template.

Description:

· The enzyme is purified from a recombinant source.

· The only clone truly derived from the Avian Myeloblastosis Virus.

· Maintains the RNA- and DNA-dependent DNA polymerase and RNase H activities.

· RNase H activity can be regulated over a wide range of temperatures.

· Expressed as a very stable, highly active polymer.

· Remarkably robust in cDNA synthesis and RT-PCR.

· Capable of synthesizing cDNA over a wide range of temperatures.

· Ideal for use in RT-PCR, cDNA libraries, RAMP™, NASBA™ and dideoxy-DNA sequencing (1,2,3).

Unit Definition: One unit is the amount of enzyme required to incorporate 1 nmol of dTTP into acid-insoluble form in 10 min at 37^oC (4).

Storage Conditions: Store at –20^oC.

Storage Buffer: 200 mM potassium phosphate (pH 7.2), 2% (v/v) Triton™X-100 and 50% (v/v) glycerol.

Assay Conditions: 50 mM Tris-HCl (pH 8.3 at 22^oC), 6 mM MgCl₂, 40 mM KCl,1 mM dithiothreitol, 0.2 mM poly(rA)·(dT)₅₀, 0.5 mM [³H]dTTP in a reaction volume of 50 µl.

Quality Control: All preparations are assayed for contaminating endonuclease and exonuclease and nonspecific RNase and single- and double- stranded DNase activities.

References:

1. Goodman, H.M. and MacDonald, R.J. (1979) Methods Enzymol. 68, 75-90.

2. Naylor, L.H. and van de Sande, J.H. (1986) Nucleic Acids Res. 14, 5939.

3. Zagursky, R.J., Baumeister, K., Lomax, N. and Berman, M.L. (1985) Gene Anal. Techn. 2, 89-94.

4. Houts, G.E., Masakau, M., Ellis, C., Beard, D. and Beard, J.W. (1979) J. Virol. 29, 517-522.