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mRNA Decapping Enzyme

Catalog # Concentration Size List Price Quantity Your Price
M0608S 100000 units/ml 2000 units $157.00
$141.30
Catalog # Size List Price Your Price
M0608S 2000 units $157.00
$141.30
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

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
 
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
mRNA Decapping Enzyme from S.pombe is expressed as a His-tagged fusion in E. coli.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  mRNA Decapping Enzyme M0608SVIAL -20 1 x 0.02 ml 100,000 units/ml
  mRNA Decapping Enzyme Reaction Buffer B0608SVIAL -20 1 x 1 ml 10 X
An exception occurred during the operation, making the result invalid. Check InnerException for exception details.

Companion Products
References
  • Paquette, DR., et. al (2018). RNA. 24 (2), 251-257.
  • Grudzien-Nogalska, E., and Kiledjian M (2017). Wiley Interdiscip Rev RNA. 8, e1379.
Quality Control Assay
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 Data Sheets
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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.This product is for in vitro research use only.

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