AMV Reverse Transcriptase

The T size of this product has been discontinued.  The S and L size will still be available.
unique buffer Icon_IncTemp42 heat inactivation
Catalog #SizeConcentrationPriceQtyAdd to Cart
M0277S200 units10,000 units/ml$69.00Add to Cart
M0277L1,000 units10,000 units/ml$276.00Add to Cart
  
Categories:
cDNA Synthesis & Reverse Transcriptases,
cDNA Synthesis & Reverse Transcriptases,
Reverse Transcriptases & RT-PCR Products
Applications:
cDNA Synthesis,
cDNA Synthesis,
cDNA Synthesis & RT-PCR

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).

Highlights

  • Synthesizes cDNA from single-stranded RNA

Product Source

Avian Myeloblastosis Vi­rus (AMV)

Reagents Supplied

The following reagents are supplied with this product:

Store at (°C)Concentration
AMV Reverse Transcriptase Reaction Buffer10X

Advantages and Features

Applications

  • cDNA Synthesis
  • RNA Sequencing
  • RT-PCR

Properties and 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 Temperature

-20°C

Storage Conditions

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 min

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.

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 Laboratory Manual. (2nd ed.), 8-64. Cold Spring Harbor Laboratory Press.
  3. Kacian, D.L. (1977). Meth. Virol.. 6, 143.

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)

Datacards

The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.

Selection Charts

Usage Guidelines & Tips

Interactive Tools

Quality Control

Quality Control Assays

The following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page. Further information regarding NEB product quality can be found here.
  • Endonuclease Activity (Nicking):
    The product is tested in a reaction containing a supercoiled DNA substrate. After incubation for 4 hours the percent converted to the nicked form is determined by agarose gel electrophoresis.
  • Exonuclease Activity (Radioactivity Release):
    The product is tested in a reaction containing a radiolabeled mixture of single and double-stranded DNA. After incubation for 4 hours the exonuclease activity is determined by the % release of radioactive nucleotides.
  • RNase Activity (2 Hour Digestion):
    The product is tested in a reaction containing a RNA substrate.  After incubation for 2 hour there is no detectable degradation of the RNA substrate as determined by gel electrophoresis.

Certificate of Analysis

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

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]

Safety Data Sheet

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

Datacards

The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.