Product Class: Other

RNA 5' Pyrophosphohydrolase (RppH)
NEB2 cloned at NEB recombinant 37

Product Introduction

RNA 5´ Pyrophosphohydrolase (RppH) removes pyrophosphate from the 5´ end of triphosphorylated RNA to leave a 5´ monophosphate RNA.

  • Isolated from a recombinant source
  • Supplied with 10X Reaction Buffer
  • Tested for the absence of endonucleases, exonucleases, RNases and phosphatases
Catalog # Size Concentration
M0356S 200.0 units 5000 units/ml

Product Information

Description

The bacterial RNA 5´ Pyrophosphohydrolase (RppH) removes pyrophosphate from the 5´ end of triphosphorylated RNA to leave a 5´ monophosphate RNA (1). The RppH protein was also known as NudH/YgdP which can split Ap5A to ADP and ATP (2).

Product Source

An E. coli strain containing a clone of the E. coli RppH gene.
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

Properties & Usage

Unit Definition

One unit is the amount of enzyme that converts 1 µg 300 mer RNA transcript into a XRN-1 digestible RNA in 30 minutes at 37°C.

Reaction Conditions

1X NEBuffer™ 2
Incubate at 37°C

1X NEBuffer™ 2
50 mM NaCl
10 mM Tris-HCl
10 mM MgCl2
1 mM DTT
(pH 7.9 @ 25°C)

Storage Buffer

20 mM Tris-HCl
200 mM NaCl
1 mM DTT
0.1 mM EDTA
50% Glycerol
0.01% Triton® X-100
pH 7.5 @ 25°C

Product Notes

  1. Since RppH is active in the presence of Mg2+, it can be incubated with RNA Ligase or XRN-1.

References

  1. Deana, A. et al. (2008). Nature. 451, 355-358.
  2. Bessman, M.J. et al. (2001). JBC. 276, 37834.
  3. Neri F, Rapelli S, Krepelova A, Incarnato D, Parlato C, Basile G, et al. (2017). Intragenic DNA Methylation prevents spurious transcription initiation. Nature. 543, 72–7.
  4. Hetzel, Jonathan et al. (2016). Nascent RNA Sequencing Reveals Distinct Features in Plant Transcription. Proceeding of the National Academy of Sciences. 113.43, 12316–12321.

Protocols, Manuals & Usage

Protocols

  1. Decapping eukaryotic mRNA with RppH (NEB #M0356)
  2. Cappable-seq for prokaryotic transcription start site (TSS) determination
  3. Removal of pyrophosphate from 5 triphosphorylated RNA using RppH (NEB #M0356)

FAQs & Troubleshooting

FAQs

  1. Can RppH replace Tobacco Acid Pyrophosphatase (TAP) as a decapping enzyme?

Tech Tips

We strongly recommend wearing gloves, using nuclease-free tubes and reagents, and thoroughly cleaning pipettes and bench surfaces to avoid RNase contamination.

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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