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RtcB Ligase

Catalog # Concentration Size List Price Quantity Your Price
M0458S 15 µM 25 reactions $122.00
$109.80
Catalog # Size List Price Your Price
M0458S 25 reactions $122.00
$109.80
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca
  • This is an Enzyme for Innovation (EFI). EFI is a project initiated by NEB to provide unique enzymes to the scientific community in the hopes of enabling the discovery of new and innovative applications. These enzymes have interesting properties and unique specificities.
  • RtcB Ligase from E. coli joins single stranded RNA with a 3´-phosphate or 2´,3´-cyclic phosphate to another RNA with a 5´ -hydroxyl (1,2).
  • Ligation requires both GTP and MnCl2 and proceeds through a 3´-guanylate intermediate (3).
  • For substrates with a 2´,3´-cyclic phosphate end, hydrolysis to a 3´-phosphate precedes 3´ end activation with GMP and ligation.
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Product Source
RtcB Ligase 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
  RtcB Ligase M0458SVIAL -20 1 x 0.025 ml 15 µM
  RtcB Reaction Buffer B0790AVIAL -20 1 x 0.5 ml 10 X
  MnCl2 B1761SVIAL -20 1 x 1 ml 10 X
  GTP N2080AVIAL -20 1 x 0.05 ml 10 mM
Application Features
  • Ligate ssRNA or ssDNA with a 3´-phosphate or a 2´,3´-cyclic phosphate to the 5´-OH of ssRNA
  • Circularization of ssRNA with compatible ends, as above
  • Detection of 3´-phosphorylated or 5´ -OH RNA using labeled probes

Properties & Usage

Reaction Conditions

1X RtcB Reaction Buffer
Supplement with 0.1 mM GTP and 1 mM MnCl2
Incubate at 37°C

1X RtcB Reaction Buffer
50 mM Tris-HCl
75 mM KCl
3 mM MgCl2
10 mM DTT
(pH 8.3 @ 25°C)

Storage Buffer

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



Notes
  • For difficult substrates, addition of PEG 8000 up to 15% can improve ligation yields.
References
  • Naoko, T., Shuman, S. (2011). J. Biol. Chem. 286, 7727-7731.
  • Naoko, T. et al. (2011). J. Biol. Chem. 286, 43134-43143.
  • Chakravarty, A. K. et al (2012). Proc. Natl. Acad. Sci. USA. 109, 6072-6077.
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
Legal And Disclaimer

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 busdev@neb.com.

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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