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| Code No. | Description | Unit | Conc. |
| RTA-001 | AMV Reverse Transcriptase | 300U | 10U/µl |
| RTA-002 | AMV Reverse Transcriptase | 1,000U | 10U/µl |
| RTA-003 | AMV Reverse Transcriptase (highly Concentrated) | 600U | 20-25U/µl |
| RTB-001 | Reverse Transcription 10 x Buffer | 1.4ml | |
| RTM-001 | Magnesium Chloride Solution | 1.5ml | 25mM |
| RTS-001 | Set of dATP, dCTP, dGTP, dTTP | 40µmol each | 100mM |
Description Avian Myeloblastosis Virus Reverse Transcriptase (AMV RT) catalyzes the polymerization of DNA using template DNA, RNA or RNA:DNA hybrids (1). 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 Available at High Concentration: Cat.# RTA-003 contains 600 units of AMV RT at 20–25u/μl.
Provided with 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 (2). RT-PCR(a). 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 NormTaq DNA Polymerase or PCR Master Mix are used, up to 25μl of an RT reaction can be added per 50μl PCR.
Storage Conditions Store at –20°C.
Storage Buffer 200mM potassium phosphate (pH 7.2 at 4°C), 0.2% Triton X-100, 2mM DTT and 50% gycerol.
Unit Definition One unit is defined as the amount of enzyme required to catalyze the transfer of 1nmol of dTTP into acid-insoluble form in 10 minutes at 37°C. The reaction conditions are: 50mM Tris-HCl (pH 8.3 at 25°C), 40mM KCl, 8.75mM MgCl2, 10mM DTT, 0.1mg/ml acetylated BSA, 1mM radiolabeled dTTP and 0.25mM poly(A):oligo(dT).
Quality Control Tests Activity, SDS-PAGE/purity, DNase, RNase, endonuclease, RT-PCR, first-strand cDNA synthesis.
Source Purified Avian Myeloblastosis Virus particles.
References Kacian, D.L. (1977) Meth. Virol. 6, 143. Mierendorf, R.C. and Pfeffer, D. (1987) Meth. Enzymol. 152, 563–6.
