Product Class: Other

M-MuLV Reverse Transcriptase
NEBU recombinant 42 65 Heat

Product Introduction

M-MuLV Reverse Transcriptase synthesizes a complementary DNA strand initiating from a primer using either RNA (cDNA synthesis) or single-stranded DNA as a template.

  • Isolated from a recombinant source
  • Supplied with 10X Reaction Buffer
  • Tested for the absence of DNases and RNases
Catalog # Size Concentration
M0253S 10000.0 units 200000 units/ml
M0253L 50000.0 units 200000 units/ml

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Product Information

Description

Moloney Murine Leukemia Virus (M-MuLV, MMLV) Reverse Transcriptase is an RNA-directed DNA polymerase. This enzyme can synthesize a complementary DNA strand initiating from a primer using either RNA (cDNA synthesis) or single-stranded DNA as a template. (1-4). M-MuLV Reverse Transcriptase lacks 3´ → 5´ exonuclease activity.

Product Source

The gene encoding M-MuLV Reverse Transcriptase is expressed in E. coli in a vector that results in 16 additional amino acids at the N-terminus and 13 amino acids at the C-terminus. This construct results in a fully functional Reverse Transcriptase protein with a functional RNase H domain.
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:
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
  • M0253S     -20    
      M-MuLV Reverse Transcriptase M0253SVIAL -20 1 x 0.05 ml 200,000 units/ml
      M-MuLV Reverse Transcriptase Reaction Buffer B0253SVIAL -20 1 x 1.5 ml 10 X
  • M0253L     -20    
      M-MuLV Reverse Transcriptase M0253LVIAL -20 1 x 0.25 ml 200,000 units/ml
      M-MuLV Reverse Transcriptase Reaction Buffer B0253SVIAL -20 1 x 1.5 ml 10 X

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme that will incorporate 1 nmol of dTTP into acid-insoluble material in a total reaction volume of 50 μl in 10 minutes at 37°C using poly(rA).oligo(dT) as template primer with 50 mM Tris-HCI (pH 8.3), 6 mM MgCl2, 10 mM dithiothreitol, 0.5 mM [3H]-dTTP and 0.4 mM poly(rA).oligo(dT)12-18.

Reaction Conditions

1X M-MuLV Reverse Transcriptase Reaction Buffer
Incubate at 42°C

1X M-MuLV Reverse Transcriptase Reaction Buffer
50 mM Tris-HCl
75 mM KCl
3 mM MgCl2
10 mM DTT
(pH 8.3 @ 25°C)

Storage Buffer

50 mM Tris-HCl
150 mM NaCl
1 mM DTT
0.1 mM EDTA
50% Glycerol
0.1% NP-40
pH 7.6 @ 25°C

Heat Inactivation

65°C for 20 minutes

Molecular Weight

Theoretical: 78000 daltons

References

  1. Verma, I.M. (1975). J.Virol.. 15, 843-854.
  2. Gerard, G.F. and Grandgenett, D.P. (1975). J.Virol.. 15, 785-797.
  3. Roth, M.J., Tanese, N. and Goff, S.P. (1985). J. Biol.Chem.. 260, 9326-9335.
  4. Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual(2nd Ed.). 5.52-5.55, 8.11-8.17.

Protocols, Manuals & Usage

Protocols

  1. First Strand cDNA Synthesis (Quick Protocol) (NEB #M0253)
  2. First Strand cDNA Synthesis (Standard Protocol) (NEB #M0253)
  3. First Strand cDNA Synthesis (No-RT Negative Control Reaction) (NEB #M0253)

Usage & Guidelines

Tools & Resources

Selection Charts

FAQs & Troubleshooting

FAQs

  1. Can M-MuLV Reverse Transcriptase be used in other NEBuffers?
  2. Does M-MuLV Reverse Transcriptase require DTT?
  3. Can the M-MuLV Reverse Transcriptase transcribe through RNA-DNA junctions?
  4. How can the length of the product generated by M-MuLV Reverse Transcriptase be increased?
  5. How can the yield be improved when using M-MuLV Reverse Transcriptase?
  6. What are some of the reasons for M-MuLV Reverse Transcriptase reaction failure?
  7. Is M-MuLV Reverse Transcriptase active at temperatures higher than 42°C?
  8. Can DNA be used as a template for M-MuLV Reverse Transcriptase?
  9. Are NEB DNA Polymerases supplied with dNTPs?

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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