mRNA Decapping Enzyme
NEBU No

mRNA Decapping Enzyme decaps m7G-capped mRNA producing a 5′ monophosphate end.

  • Efficient replacement for Tobacco Acid Pyrophosphatase
  • Cap0 and Cap1 are removed with equal efficiency
  • Suitable for 5′ RLM-RACE and RNA-seq
  • 1 µl will decap 20 µg of a 1.7 kb mRNA in 15 minutes
 
Catalog # Concentration Size List Price Your Price Quantity
M0608S 100,000 units/ml 2,000 units $95.00
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  • Product Information
    mRNA Decapping Enzyme catalyzes the removal of 7-methylguanosine cap (m7G) from 5´ end of mRNA, producing 5′ monophosphate and releasing m7GDP(1). mRNA Decapping Enzyme is capable of decapping mRNAs of various lengths and removes both Cap0 and Cap1 structures with similar efficiency. mRNA Decapping Enzyme also converts 5′ triphosphate ends to 5′ monophosphate, albeit with reduced efficiency. 5′ monophosphorylated RNA can be exploited in a variety of downstream applications, including 5′ RNA Ligase-mediated RACE, RNA-seq, and 5′-> 3′ exonuclease digestion.

    Comparison of mRNA Decapping Enzyme and Tobacco Acid Pyrophosphatase Decapping Activity.



    (Panel A) A 500 nM solution of 25 nucleotide 5′ capped RNA was treated with either mRNA Decapping Enzyme (MDE) or competitor Tobacco Acid Pyrophosphatase (TAP). Both reactions were carried out in the same reaction volume and using each enzyme’s appropriate reaction buffer. While the concentration of a competitor’s TAP used is unknown, this comparison reveals 0.62 units MDE to have a similar level of activity relative to 1 µl of TAP. Decapping reaction products were analyzed by capillary electrophoresis. Note that in the negative control reaction there is a contaminating amount of triphosphorylated RNA of ~10% in the synthetic substrate used.
    (Panel B) To test the impact of introducing length and termini heterogeneity into the reaction, 500 ng of total RNA for Jurkat cells was added while maintaining all other conditions unchanged. Decapping by MDE was unaffected, whereas TAP activity was markedly diminished, indicating that MDE is the more robust decapping enzyme under these test conditions.
     

     

    Product Source

    An E. coli strain that carries a plasmid encoding the mRNA Decapping Enzyme.

    Reagents Supplied

    The following reagents are supplied with this product:

    Store at (°C) Concentration
    mRNA Decapping Enzyme Reaction Buffer -20
    Product Categories:
    RNA Modification Products,
    RNA Reagents Products
    Applications:
    RNA Modification,
    RNA Analysis
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