Product Class: Other

yDcpS
NEBU recombinant 37 70 Heat

Product Introduction

yDcpS is a decapping enzyme that hydrolyzes m7G capped mRNA, producing diphosphorylated 5′ ends. yDcpS can decap mRNAs of various lengths and the diphosphorylated 5′ end can be recapped directly with Vaccinia Capping Enzyme.

  • Provided at high enzyme concentration to facilitate complete decapping
  • Active on capped RNA of various sizes (tested on in vitro capped transcripts from 25 nt to 1400 nt long)
  • Decapped RNA 5′ ends can be recapped with Vaccinia Capping System (NEB #M2080)
  • Supports tagging of mRNA with desthiobiotin-labeled GTP
Catalog # Size Concentration
M0463S 4000.0 units 200000 units/ml

Product Information

Description

yDcpS is the scavenging decapping enzyme from S. cerevisiae that hydrolyzes the phosphodiester bond between the gamma and beta phosphates of m7G-capped mRNA, producing diphosphorylated 5′ ends and m7GMP.1,2 yDcpS is capable of decapping mRNAs of various lengths, decaps Cap0 and Cap1 with equal efficiency, and the diphosphorylated 5′ end it produces is suitable for recapping with Vaccinia Capping Enzyme.3,4

Product Source

A His-tagged engineered variant of E. coli that carries the S. cerevisiae gene DCS1 (encoding yDcpS) on a plasmid.
This product is related to the following categories:
RNA Modification

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • M0463S     -20    
      yDcpS M0463SVIAL -20 1 x 0.02 ml 200,000 units/ml
      yDcpS Reaction Buffer B0463AVIAL -20 1 x 1 ml 10 X

Properties & Usage

Unit Definition

One unit is defined as the amount of yDcpS required to convert 50% of a 500 nM solution of the following 25-mer m7G-capped RNA to a 5’-diphosphorylated form in a total reaction volume of 20 µl in 1 hour at 37˚C: 5’-[m7Gppp]rGrUrArGrArArCrUrUrCrGrUrCrGrArGrUrArCrGrCrUrCrArA[3-6FAM]-3’

Reaction Conditions

1X yDcpS Reaction Buffer
Incubate at 37°C

1X yDcpS Reaction Buffer
10 mM Bis-Tris-HCl
1 mM EDTA
(pH 6.5 @ 25°C)

Storage Buffer

300 mM NaCl
10 mM Tris-HCl
0.1 mM EDTA
1 mM DTT
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation

70°C

Specific Activity

20,000 units/mg

Unit Assay Conditions

1X yDcpS Reaction Buffer with 500 nM m7G-capped 25-mer 3´-FAM RNA. After incubation at 37°C for 1 hour, yDcpS is inactivated at 70˚C for 15 minutes and decapping is measured by capillary electrophoresis.

Features

  • mRNA decapping enabling recapping with tagged-GTP analogs
  • Biotinylation of 5′ ends of primary transcripts
  • Recappable-seq

References

  1. Wang, X. and Kiledjian, M. (2001). Cell. 107 (6), 751-62.
  2. Liu, H., Rodgers, N.D., Jiao, X., and Kiledjian, M. (2002). EMBO J.. 21(17), 4699-708.
  3. Wulf MG, Buswell J, Chan SH, Dai N, Marks K, Martin ER, Tzertzinis G, Whipple JM, Corrêa IR Jr, Schildkraut I. (2019). The yeast scavenger decapping enzyme DcpS and its application for in vitro RNA recapping. Sci Rep. Jun 13; 9 (1), 8594.
  4. Bo Yan, George Tzertzinis, Ira Schildkraut, Laurence Ettwiller ReCappable Seq: Comprehensive Determination of Transcription Start Sites derived from all RNA polymerases. bioRxiv. 696559.

Protocols, Manuals & Usage

Protocols

  1. Protocol for decapping of total RNA (NEB #M0463)

FAQs & Troubleshooting

FAQs

  1. Is yDcpS an alternative for Tobacco Acid Pyrophosphotase (TAP)?
  2. In the literature DcpS is reported to have no activity on transcripts longer that 25 nt. Is yDcpS from NEB active on longer transcripts?
  3. Is yDcpS active in Capping Buffer supplied with the Vaccinia Capping System (NEB #M2080)?
  4. What is the target specificity of yDcpS?
  5. Is yDcpS activity sensitive to RNA secondary structure?
  6. What are the preferred buffer conditions for yDcpS?
  7. Is yDcpS active at higher temperatures than 37˚C?

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 [email protected].

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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