M-MuLV Reverse Transcriptase

recombinant unique buffer heat inactivation
Catalog #SizeConcentrationPriceQtyAdd to Cart
M0253S10,000 units200,000 units/ml$68.00Add to Cart
M0253L50,000 units200,000 units/ml$272.00Add 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

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.

Highlights

  • Isolated from a recombinant source
  • Synthesizes cDNA from single-stranded RNA or DNA

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.

Reagents Supplied

The following reagents are supplied with this product:

Store at (°C)Concentration
M-MuLV Reverse Transcriptase Reaction Buffer-2010X

Properties and 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 Temperature

-20°C

Storage Conditions

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 min

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.

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. Are NEB DNA Polymerases supplied with dNTPs?
  5. How can the length of the product generated by M-MuLV Reverse Transcriptase be increased?
  6. How can the yield be improved when using M-MuLV Reverse Transcriptase?
  7. What are some of the reasons for M-MuLV Reverse Transcriptase reaction failure?
  8. Is M-MuLV Reverse Transcriptase active at temperatures higher than 42°C?
  9. Can DNA be used as a template for M-MuLV Reverse Transcriptase?

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)

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 Assurance Statement

  • M-MuLV Reverse Transcriptase is tested for its ability to synthesize full length cDNAs from crude or purified RNA templates. Purified free of detectable levels of RNase, endonuclease and exonuclease activities.

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.
  • 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.
  • Non-Specific DNase Activity (16 hour):
    The product is tested for non-specific nuclease degradation in a reaction containing a DNA substrate. After incubation for 16 hours there is no detectable degradation of the DNA substrate as determined by agarose gel electrophoresis.
  • 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.