T4 RNA Ligase 2 (dsRNA Ligase)

Product Information

• Reagents Supplied

  The following reagents are supplied with this product:

  NEB https://www.neb.com/en/#	Component Name	Component https://www.neb.com/en/#	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.

• Advantages and Features

  Application Features

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

• Related Products

  Companion Products

  • T4 RNA Ligase 1 (ssRNA Ligase)
  • RNase Inhibitor, Murine

• 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 https://www.neb.com/en/#M0239)
  2. Protocol for ligation of 3′OH of RNA and 5′PO₄ of DNA in ds structure with T4 RNA Ligase 2 (NEB https://www.neb.com/en/#M0239)

Tools & Resources

• Selection Charts

  • Properties of DNA and RNA Ligases
  • RNA Ligase Selection Chart
  • Substrate-based Ligase Selection Chart

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?

Citations & Technical Literature

• Citations

  Product Citation Tool

Quality, Safety & Legal

• Quality Control Assays

  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]

  • M0239L_v1_10016831
  • M0239L_v1_10029433
  • M0239L_v1_10031202
  • M0239L_v1_10039469
  • M0239S_v1_10039471
  • M0239L_v1_10056214
  • M0239S_v1_10052770
  • M0239L_v1_10059007
  • M0239S_v1_10056216
  • M0239L_v1_10062561
  • M0239L_v1_10068405
  • M0239L_v1_10077151
  • M0239S_v1_10081143
  • M0239L_v1_10080149
  • M0239L_v1_10088324
  • M0239S_v1_10088326
  • M0239L_v1_10102475
  • M0239L_v1_10102473
  • M0239S_v1_10120087
  • M0239L_v1_10124275
  • M0239S_v1_10146850
  • M0239S_v1_10156049
  • M0239L_v1_10158106
  • M0239S_v1_10167889
  • M0239L_v1_10176627
  • M0239L_v1_10178719
  • M0239L_v1_10181512
  • M0239S_v1_10183839
  • M0239L_v1_10200635
  • M0239S_L_v1_0041802
  • M0239S_v1_10220914
  • M0239L_v1_10226275
  • M0239S_v1_10246121
  • M0239S_v1_10272939

• Safety Data Sheets

  The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

  • T4 RNA Ligase 2 (dsRNA Ligase)
  • T4 Rnl2 Reaction Buffer

• 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.