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Home > Products > Polymerases > Reverse Transcriptases and RNA Polymerases > AMV Reverse Transcriptase AMV Reverse Transcriptase Catalog # Size Concentration Price Qty   M0277L 1,000 units 10,000 units/ml $240.00 M0277S 200 units 10,000 units/ml $60.00 M0277T 500 units 25,000 units/ml $135.00 Prices are in US dollars and valid only for US orders. Download: MSDS PDF Synthesizes cDNA from single-stranded RNA Description: Avian Myeloblastosis Vi­rus (AMV) Reverse Transcriptase is an RNA-directed DNA polymerase. This en­zyme can synthesize a complementary DNA strand initiating from a primer using RNA (cDNA synthesis). (1-3) Source: Avian Myeloblastosis Vi­rus (AMV) Applications: cDNA Synthesis RNA Sequencing RT-PCR Reagents Supplied: AMV Reverse Transcriptase Reaction Buffer (10X) Reaction & Storage Conditions Reaction Conditions: 1X AMV Reverse Transcriptase Reaction Buffer Supplemented with Incubate at 37°C. 1X AMV Reverse Transcriptase Reaction Buffer: 50 mM Tris-HCl 75 mM potassium acetate 8 mM Magnesium Acetate 10 mM DTT pH 8.3 @ 25°C Unit Definition: One unit is defined as the amount of enzyme required to incorporate 1 nmol of dTMP into an acid-insoluble form in 10 minutes at 37°C using poly(rA)-oligo(dT) as template primer.  Unit Assay Conditions: 75 mM potassium acetate, 50 mM Tris-HCl (pH 8.3) 8 mM magenesium acetate, 0.5 mM [3H]-dTTP, 0.2 mM poly(rA)-oligo(dT)12-18. Concentration: 10,000 units/ml and 25,000 units/ml Storage Conditions: 0.2 M potassium phosphate 2 mM Dithiothreitol 50% Glycerol 0.2% Triton X-100 pH 7.2 @ 25°C Storage Temperature: -20°C Notes General notes: The yield and size of cDNA transcript increases with increasing amounts of RT. Storage: Once thawed, store at -20°C. Repeated freeze thaw cycle will inactivate the enzyme. Aliquots can be stored for longer periods at -70°C. Usage notes: Reaction Conditions: Incubate at 37°C or 42°C. Quality Control for Current Lot Quality control values for a specific lot can be found on the datacard which accompanies each product. 16-Hour Incubation: A 50 μl reaction containing 1 μg of ΦX174 RF I DNA (HaeIII digest) and 30 units of AMV Reverse Transcriptase incubated for 16 hours at 37ºC resulted in a DNA pattern free of detectable nuclease degradation as determined by agarose gel electrophoresis. Exonuclease Activity: Incubation of a 50 μl reaction containing 30 units of AMV Reverse Transcriptase with 1 μg of a mixture of single and double-stranded [3H] E. coli DNA (205 cpm/μg) for 4 hours at 37ºC released < 0.2% of the total radioactivity. RNase Assay: Incubation of a 50 μl reaction containing 30 units of AMV Reverse Transcriptase with 3 μg of dsRNA ladder for 1 hour at 37ºC resulted in no detectable degradation of the RNA as determined by agarose gel electrophoresis. References Kacian, D.L. (1977) Meth. Virol., 6, 143. Krug, M.S. et al. (1987) Meth. Enzymol., 152, 316-325. Sambrook.J., Fritsch, E.F. and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, (2nd ed.), 8-64. Cold Spring Harbor: Cold Spring Harbor Labo.