Product Class: Other

AMV Reverse Transcriptase
NEBU 42 85 Heat

The T size of this product has been discontinued.  The S and L size will still be available.

Product Introduction

  • DNA synthesis from either RNA or ssDNA
  • Ideal for RT-PCR, cDNA synthesis, and RNA sequencing
  • 10X reaction buffer creates optimal reaction conditions for robust first and second-strand cDNA synthesis
Catalog # Size Concentration
M0277S 200.0 units 10000 units/ml
M0277L 1000.0 units 10000 units/ml

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Product Information

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) or single-stranded DNA as a template (1-3).

Product Source

Insect cells (Sf21) infected with baculovirus containing the pol gene of AMV.
This product is related to the following categories:
RT-PCR Products,
cDNA Synthesis & Reverse Transcriptases Products,
This product can be used in the following applications:
Transcription-Mediated and NASBA Amplification,
cDNA Synthesis,
Reverse Transcription (cDNA Synthesis),
RT-PCR & cDNA Synthesis,
PCR

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • M0277S     -20    
      AMV Reverse Transcriptase M0277SVIAL -20 1 x 0.02 ml 10,000 units/ml
      AMV Reverse Transcriptase Reaction Buffer B0277AVIAL -20 1 x 1.5 ml 10 X
  • M0277L     -20    
      AMV Reverse Transcriptase M0277LVIAL -20 1 x 0.1 ml 10,000 units/ml
      AMV Reverse Transcriptase Reaction Buffer B0277AVIAL -20 1 x 1.5 ml 10 X

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to incorporate 1 nmol of dTTP into an acid-insoluble form in 10 minutes at 37°C using poly(rA)-oligo(dT) as template primer.

Reaction Conditions

1X AMV Reverse Transcriptase Reaction Buffer
Incubate at 42°C

1X AMV Reverse Transcriptase Reaction Buffer
50 mM Tris-acetate
75 mM Potassium Acetate
8 mM Magnesium Acetate
10 mM DTT
(pH 8.3 @ 25°C)

Storage Buffer

200 mM potassium phosphate
2 mM DTT
50% Glycerol
0.2% Triton® X-100
pH 7.2 @ 25°C

Heat Inactivation

85°C for 5 minutes

Unit Assay Conditions

75 mM potassium acetate, 50 mM Tris-HCl (pH 8.3) 8 mM magnesium acetate, 0.5 mM [3H]-dTTP, 0.2 mM poly(rA)•oligo(dT)12–18 and 10 mM DTT.

Application Features

  • cDNA Synthesis
  • RNA Sequencing
  • RT-PCR

Product Notes

  1. Reaction Conditions: Incubate at 37-52°C.
  2. The yield and size of cDNA transcript increases with increasing amounts of RT.
  3. Storage: Once thawed, store at -20°C. Repeated freeze thaw cycles will inactivate the enzyme. Aliquots can be stored for longer periods at -70°C.

References

  1. Krug, M.S. et al. (1987). Meth.Enzymol. 152,
  2. Sambrook.J., Fritsch, E.F. and Maniatis, T. (1989). Molecular Cloning: A LaboratoryManual. (2nd ed.), 8-64. Cold Spring Harbor Laboratory Press.
  3. Kacian, D.L. (1977). Meth.Virol.. 6, 143.

Protocols, Manuals & Usage

Protocols

  1. First Strand cDNA Synthesis (Quick Protocol) (NEB #M0277)
  2. First Strand cDNA Synthesis (Standard Protocol) (NEB #M0277)
  3. First Strand cDNA Synthesis (No-RT Negative Control Reaction) (NEB #M0277)

Usage & Guidelines

Tools & Resources

Selection Charts

Quality, Safety & Legal

Quality Assurance Statement

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

Specifications

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

Certificate Of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

Safety DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

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