AMV Reverse Transcriptase:Part No.M510AM510FM900A(High Conc.) 600Size (units)3001,000AMV Reverse Transcriptase 5X Reaction Buffer (M515A): The AMV Reverse Transcriptase 5X Reaction Buffer suppliedwith this enzyme has a composition of 250mM Tris-HCl (pH 8.3 @ 25°C), 250mM KCl, 50mM MgCl2, 2.5mM spermidine and50mM DTT.Enzyme Storage Buffer: AMV Reverse Transcriptase (AMV-RT) is supplied in 200mM potassium phosphate (pH 7.2 @4°C), 0.2% Triton®X-100, 2mM DTT and 50% glycerol.Source: Purified from avian myeloblastosis virus particles.Storage Conditions: Store at –20°C. Avoid multiple freeze-thaw cycles and exposure to frequent temperature changes.See the expiration date on the Product Information Label.Unit Definition: One unit is defined as the amount of enzyme required to catalyze the transfer of 1nmol of deoxynucleotideinto acid-precipitable material in 10 minutes at 37°C. The reaction conditions are: 50mM Tris-HCl (pH 8.3), 40mM KCl,8.75mM MgCl2, 10mM DTT, 0.1mg/ml acetylated BSA, 1mM radiolabeled dTTP and 0.25mM poly(A):oligo(dT). See the unitconcentration on the Product Information Label.Usage Notes:1. The AMV Reverse Transcriptase 5X Reaction Buffer is intended for use in standard first-strand cDNA synthesis reactions. Nodeoxynucleotides are in the buffer; therefore, this buffer must not be substituted for the Promega RiboClone®AMV RT First-Strand 5X Buffer (Part# C121A), a component of the Universal RiboClone®cDNA Synthesis System (Cat.# C4360), whichdoes have dNTPs. The Access RT-PCR System (Cat.# A1250) utilizes AMV Reverse Transcriptase and Tfl DNA Polymerase toprovide a combined reverse transcription and PCR without intermediate handling. The reaction buffer provided in the AccessRT-PCR System is not the same as the 5X Reaction Buffer provided with AMV-RT. The two buffers are not interchangeable.2. The formulation of AMV Reverse Transcriptase 5X Reaction Buffer is not compatible with M-MLV Reverse Transcriptase.3. Up to 10µl of an RT reaction containing AMV-RT and the supplied AMV Reverse Transcriptase Reaction Buffer can be addedto PCR amplification reactions that use Taq DNA Polymerase. If GoTaq®DNA Polymerase (Cat.# M3001) or PCR Master Mix(Cat.# M7501) are used, up to 25µl of the RT reaction can be added to a 50µl PCR.Quality Control AssaysActivity AssayFirst-Strand cDNA Synthesis: First-strand cDNA, of a 1.2kb Control RNA (from Cat.# C4360), is synthesized using 30units of AMV Reverse Transcriptase per microgram of template, an oligo(dT) primer and a radiolabeled dNTP. The minimumspecification is the conversion of >12% of mRNA to cDNA. Full-length cDNA must be observed by gel electrophoresis andautoradiography.Contaminant ActivityEndonuclease Assay: To test for endonuclease activity, 1µg of Type I supercoiled plasmid DNA is incubated with 25 units ofAMV Reverse Transcriptase in 50mM Tris (pH 8.3), 40mM KCl, 7mM MgCl2, 10mM DTT for one hour at 37°C. Following incubation, the supercoiled DNA is visualized on an ethidium bromide-stained agarose gel to verify the absence of visible nicking or cutting.DNase and RNase Assay: To test for nuclease activity, 50ng of radiolabeled DNA or radiolabeled RNA is incubated with 25 units of AMV Reverse Transcriptase in 4mM Tris (pH 8.3), 3.2mM KCl, 0.56mM MgCl2, 0.8mM DTT for one hour at 37°C,and the release of radiolabeled nucleotides is monitored by scintillation counting of TCA-soluble material. Minimum passing specification is <1% release for DNase and <3% release for RNase.Physical Purity: AMV Reverse Transcriptase is a 170kDa heterodimer with an α-subunit of 65kDa and a β-subunit of 94kDa.The purity is >80% in 2 bands (2 subunits) as judged by SDS-polyacrylamide gels with Coomassie®blue staining.Promega Corporation2800 Woods Hollow RoadMadison, WI 53711-5399TelephoneToll FreeFaxInternetUSA608-274-4330800-356-9526608-277-2516www.promega.comPRODUCT USE LIMITATIONS, WARRANTY, DISCLAIMERPromega manufactures products for a number ofintended uses. Please refer to the product label for theintended use statements for specific products.Promega products contain chemicals which may beharmful if misused. Due care should be exercised withall Promega products to prevent direct human contact.Each Promega product is shipped with documentationstating specifications and other technical information.Promega products are warranted to meet or exceed thestated specifications. Promega's sole obligation and thecustomer's sole remedy is limited to replacement ofproducts free of charge in the event products fail to per-form as warranted. Promega makes no other warrantyof any kind whatsoever, and SPECIFICALLY DIS-CLAIMS AND EXCLUDES ALL OTHER WARRANTIESOF ANY KIND OR NATURE WHATSOEVER, DIRECTLYOR INDIRECTLY, EXPRESS OR IMPLIED, INCLUDING,WITHOUT LIMITATION, AS TO THE SUITABILITY, PRO-DUCTIVITY, DURABILITY, FITNESS FOR A PARTICU-LAR PURPOSE OR USE, MERCHANTABILITY, CONDI-TION, OR ANY OTHER MATTER WITH RESPECT TOPROMEGA PRODUCTS. In no event shall Promega beliable for claims for any other damages, whether direct,incidental, foreseeable, consequential, or special(including but not limited to loss of use, revenue orprofit), whether based upon warranty, contract, tort(including negligence) or strict liability arising in con-nection with the sale or the failure of Promega productsto perform in accordance with the stated specifications.© 1997–2005, 2011 Promega Corporation. All RightsReserved.GoTaq, RiboClone and RNasin are trademarks ofPromega Corporation and are registered with the U.S.Patent and Trademark Office.Coomassie is a registered trademark of ImperialChemical Industries. Triton is a registered trademarkof Union Carbide Chemicals and Plastics Co., Inc.All specifications are subject to change without priornotice.Product claims are subject to change. Please contactPromega Technical Services or access the Promegaonline catalog for the most up-to-date information onPromega products.Part# 9PIM510Revised 9/11Part# 9PIM510Printed in USA. Revised 9/11Certificate of AnalysisSigned by:J. Stevens, Quality Assurance
