Product Class: Other

T4 RNA Ligase 2 (dsRNA Ligase)
NEBU cloned at NEB recombinant 37 80 Heat

Product Introduction

T4 RNA Ligase 2 ligates nicks in dsRNA and can also be used to ligate the 3´ OH of RNA to the 5´ phosphate of DNA in a double stranded structure.

  • Best choice for ligating nicks in dsRNA
  • Suitable for ligating 3' OH of RNA to 5' phosphate of DNA in a DNA/RNA hybrid
  • Isolated from a recombinant source
  • Supplied with 10X Reaction Buffer
  • Tested for the absence of endonucleases, exonucleases and RNases
  • Not sure which ligase to choose? Refer to our DNA and RNA Ligase Properties Chart 
Catalog # Size Concentration
M0239S 150.0 units 10000 units/ml
M0239L 750.0 units 10000 units/ml

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Product Information

Description

T4 RNA Ligase 2, also known as T4 Rnl2 (gp24.1), has both intermolecular and intramolecular RNA strand joining activity (1-3). Unlike T4 RNA Ligase 1 (NEB #M0204), T4 RNA Ligase 2 is much more active joining nicks on double stranded RNA than on joining the ends of single stranded RNA. The enzyme requires an adjacent 5´ phosphate and 3´ OH for ligation. The enzyme can also ligate the 3´ OH of RNA to the 5´ phosphate of DNA in a double stranded structure (4).

Product Source

An E. coli strain that carries the T4 RNA Ligase 2 gene.
This product is related to the following categories:

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • M0239S     -20    
      T4 RNA Ligase 2 (dsRNA Ligase) M0239SVIAL -20 1 x 0.015 ml 10,000 units/ml
      T4 Rnl2 Reaction Buffer B0239SVIAL -20 1 x 1.5 ml 10 X
  • M0239L     -20    
      T4 RNA Ligase 2 (dsRNA Ligase) M0239LVIAL -20 1 x 0.075 ml 10,000 units/ml
      T4 Rnl2 Reaction Buffer B0239SVIAL -20 1 x 1.5 ml 10 X

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to ligate 0.4 µg of an equimolar mix of a 23-mer and 17-mer RNAs in a total reaction volume of 20 µl in 30 minutes at 37°C.

Reaction Conditions

1X T4 Rnl2 Reaction Buffer
Incubate at 37°C

1X T4 Rnl2 Reaction Buffer
50 mM Tris-HCl
2 mM MgCl2
2 mM DTT
400 µM ATP
(pH 7.5 @ 25°C)

Storage Buffer

10 mM Tris-HCl
50 mM KCl
35 mM (NH4)2SO4
0.1 mM DTT
0.1 mM EDTA
50% Glycerol
pH 7.5 @ 25°C

Heat Inactivation

80°C for 5 minutes

Specific Activity

40,000 units/mg

Unit Assay Conditions

1X T4 Rnl2 Reaction Buffer and 0.4 μg of an equimolar mix of the 23-mer and 17-mer RNAs. After incubation at 37°C for 30 minutes, the ligated product is detected on a 15% polyacrylamide gel.

Application Features

  • Ligate a nick in dsRNA
  • Ligate the 3´ OH of RNA to the 5´ phosphate of DNA in a double stranded structure

References

  1. Ho, C.K. and Shuman, S. (2002). Proc. Natl. Acad. Sci. USA. 99, 12709-12714.
  2. Ho, C.K. et al. (2004). Structure. 12, 327-339.
  3. Nandakumar, J. et al. (2004). J. Biol. Chem.. 279, 31337-31347.
  4. Nandakuman, J. and Shuman, S. (2004). Mol. Cell. 16, 211-221.

Protocols, Manuals & Usage

Protocols

  1. Protocol for ligation of Nicks in dsRNA with T4 RNA Ligase 2 (NEB #M0239)
  2. Protocol for ligation of 3′OH of RNA and 5′PO4 of DNA in ds structure with T4 RNA Ligase 2 (NEB #M0239)

Tools & Resources

Selection Charts

FAQs & Troubleshooting

FAQs

  1. How can I achieve optimal ligation of a 5’ phosphorylated ssDNA oligo to the 3’ end of an ssRNA substrate with a ssDNA or ssRNA splint using T4 RNA Ligase 2?

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.

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 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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