Patent Pending
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.
First-Strand cDNA Synthesis
Description:
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The enzyme is purified from a recombinant source.
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The only clone truly derived from the Avian Myeloblastosis Virus.
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Maintains the RNA- and DNA-dependent DNA polymerase and RNase H activities.
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RNase H activity can be regulated over a wide range of temperatures.
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Expressed as a very stable, highly active polymer.
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Remarkably robust in cDNA synthesis and RT-PCR.
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Capable of synthesizing cDNA over a wide range of temperatures.
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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°C (4).
Storage Conditions: Store at –20°C.
Storage Buffer: 200 mM potassium phosphate (pH 7.2), 2% (v/v) Triton™X-100 and 50% (v/v) glycerol.
5 x Reaction Buffer: 250 mM Tris acetate (pH 8.4), 375 mM potassium acetate, 40 mM magnesium acetate and stabilizers.
Assay Conditions: 50 mM Tris-HCl (pH 8.3 at 22°C), 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:
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Goodman, H.M. and MacDonald, R.J. (1979) Methods Enzymol. 68, 75-90.
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Naylor, L.H. and van de Sande, J.H. (1986) Nucleic Acids Res. 14, 5939.
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Zagursky, R.J., Baumeister, K., Lomax, N. and Berman, M.L. (1985) Gene Anal. Techn. 2, 89-94.
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Houts, G.E., Masakau, M., Ellis, C., Beard, D. and Beard, J.W. (1979) J. Virol. 29, 517-522.