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T4 RNA Ligase 2, truncated

Catalog # Concentration Size List Price Quantity Your Price
M0242L 200000 units/ml 10000 units $499.00
$449.10
M0242S 200000 units/ml 2000 units $124.00
$111.60
Catalog # Size List Price Your Price
M0242L 10000 units $499.00
$449.10
M0242S 2000 units $124.00
$111.60
Catalog #
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*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca
  • T4 RNA Ligase 2, truncated (T4 Rnl2tr) specifically ligates the pre-adenylated 5´ end of DNA or RNA to the 3´ end of RNA.
  • The enzyme does not require ATP for ligation but does need the pre-adenylated substrate.
  • T4 Rnl2tr is expressed from a plasmid in E. coli, which encodes the first 249 amino acids of the full length T4 RNA Ligase 2.
  • Unlike the full length ligase, T4 Rnl2tr cannot ligate the phosphorylated 5´ end of RNA or DNA to the 3´ end of RNA.
  • This enzyme, also known as Rnl2 (1–249) has been used for optimized linker ligation for the cloning of microRNAs.
  • This enzyme reduces background ligation because it can only use pre-adenylated linkers.
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    Avoiding RNase Contamination
T4 RNA Ligase 2, truncated (T4 Rnl2 truncated) specifically ligates the pre-adenylated 5´ end of DNA or RNA to the 3´ end of RNA. The enzyme does not require ATP for ligation but does need the pre-adenylated substrate. T4 Rnl2 truncated is expressed from a plasmid in E. coli which encodes the first 249 amino acids of the full length T4 RNA Ligase 2. Unlike the full length ligase, T4 Rnl2 truncated is unable to adenylate the 5´ end of the substrate, and as a result it cannot ligate the phosphorylated 5´ end of RNA or DNA to the 3´ end of RNA (1-3). This enzyme, also known as Rnl2 (1-249) has been used for optimized linker ligation for the cloning of microRNAs. This enzyme reduces background ligation because it can only use adenylated primers (4-5).
Product Source
An E. coli strain that carries the cloned truncated T4 RNA Ligase 2 gene.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  T4 RNA Ligase 2, truncated M0242SVIAL -20 1 x 0.01 ml 200,000 units/ml
  T4 RNA Ligase Reaction Buffer B0216SVIAL -20 1 x 1.5 ml 10 X
  50% PEG 8000 B1004SVIAL -20 1 x 1 ml 1 X
  T4 RNA Ligase 2, truncated M0242LVIAL -20 1 x 0.05 ml 200,000 units/ml
  T4 RNA Ligase Reaction Buffer B0216SVIAL -20 1 x 1.5 ml 10 X
  50% PEG 8000 B1004SVIAL -20 1 x 1 ml 1 X
Application Features
  • Ligate a pre-adenylated DNA or RNA sequence tag to any RNA 3´-end
  • Join a single stranded adenylated primer to small RNAs for cDNA library creation
  • Join a single stranded adenylated primer to RNA for strand-specific cDNA library construction
T4 RNA Ligase 2, truncated | New England Biolabs
Home RNA Synthesis and Modification Products T4 RNA Ligase 2, truncated

T4 RNA Ligase 2, truncated
NEBU cloned at NEB recombinant 25 65 Heat

  • T4 RNA Ligase 2, truncated (T4 Rnl2tr) specifically ligates the pre-adenylated 5´ end of DNA or RNA to the 3´ end of RNA.
  • The enzyme does not require ATP for ligation but does need the pre-adenylated substrate.
  • T4 Rnl2tr is expressed from a plasmid in E. coli, which encodes the first 249 amino acids of the full length T4 RNA Ligase 2.
  • Unlike the full length ligase, T4 Rnl2tr cannot ligate the phosphorylated 5´ end of RNA or DNA to the 3´ end of RNA.
  • This enzyme, also known as Rnl2 (1–249) has been used for optimized linker ligation for the cloning of microRNAs.
  • This enzyme reduces background ligation because it can only use pre-adenylated linkers.

Featured Video

Catalog https://www.neb.com/en/# Concentration Size
M0242S 200,000 units/ml 2,000 units
M0242L 200,000 units/ml 10,000 units
Please enter a quantity for at least one size
Need a custom/large volume order? Contact us

  • Product Information

    T4 RNA Ligase 2, truncated (T4 Rnl2 truncated) specifically ligates the pre-adenylated 5´ end of DNA or RNA to the 3´ end of RNA. The enzyme does not require ATP for ligation but does need the pre-adenylated substrate. T4 Rnl2 truncated is expressed from a plasmid in E. coli which encodes the first 249 amino acids of the full length T4 RNA Ligase 2. Unlike the full length ligase, T4 Rnl2 truncated is unable to adenylate the 5´ end of the substrate, and as a result it cannot ligate the phosphorylated 5´ end of RNA or DNA to the 3´ end of RNA (1-3). This enzyme, also known as Rnl2 (1-249) has been used for optimized linker ligation for the cloning of microRNAs. This enzyme reduces background ligation because it can only use adenylated primers (4-5).

    Product Source

    An E. coli strain that carries the cloned truncated T4 RNA Ligase 2 gene.
    This product is related to the following categories:
    RNA Ligation
    This product can be used in the following applications:
    RNA Cloning

    • 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

    • Properties & Usage

      Unit Definition

      200 units is defined as the amount of enzyme required to give 80% ligation of a 31-mer RNA to the pre-adenylated end of a 17-mer DNA Universal miRNA Cloning Linker (https://www.neb.com/en/#S1315) in a total reaction volume of 20 µl in 1 hour at 25°C. 

      5´-FAM-rArGrUrCrGrUrArGrCrCrUrUrUrArUrCrCrGrArGrArUrUrCrArGrCrArArUrA-3´ 

      5´-rAppCTGTAGGCACCATCAAT–NH2-3´ 

      Molarity = 14 μM

      Reaction Conditions

      1X T4 RNA Ligase Reaction Buffer 
      Incubate at 25°C

      1X T4 RNA Ligase Reaction Buffer 
      50 mM Tris-HCl
      10 mM MgCl2
      1 mM DTT
      (pH 7.5 @ 25°C)

      Storage Buffer

      10 mM Tris-HCl
      100 mM NaCl
      1 mM DTT
      0.1 mM EDTA
      50% Glycerol
      pH 7.5 @ 25°C

      Heat Inactivation

      65°C for 20 minutes

      Unit Assay Conditions

      1X T4 RNA Ligase Reaction Buffer supplemented to 10% (w/v) PEG MW 8000, 20 pmol of 5´-FAM labeled RNA, and 40 pmol preadenylated DNA linker. After incubation at 25°C for 1 hour, the ligated product is detected on a 15% denaturing polyacrylamide gel.

    • Advantages and Features

      Application Features

      • Ligate a pre-adenylated DNA or RNA sequence tag to any RNA 3´-end
      • Join a single stranded adenylated primer to small RNAs for cDNA library creation
      • Join a single stranded adenylated primer to RNA for strand-specific cDNA library construction

    • Related Products

    • References

      1. Ho, C.K. et al. (2004). Structure. 12, 327-339.
      2. Ho, C.K. and Shuman, S. (2002). Proc. Natl.Acad.Sci. USA. 99, 12709-12714.
      3. Nandakumar, J. et al. (2004). J. Biol. Chem. 279, 31337-31347.
      4. Aravin, A. and Tusch, T. (2005). FEBS Letters. 579, 5830-5840.
      5. Pfeffer, S. et al. (2005). Nat. Meth.. 2, 269-276.

  • Protocols, Manuals & Usage

  • Tools & Resources

  • FAQs & Troubleshooting

  • Citations & Technical Literature

    • Citations

      Product Citation Tool

  • Quality, Safety & Legal

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References
  • Ho, C.K. et al. (2004). Structure. 12, 327-339.
  • Ho, C.K. and Shuman, S. (2002). Proc. Natl.Acad.Sci. USA. 99, 12709-12714.
  • Nandakumar, J. et al. (2004). J. Biol. Chem. 279, 31337-31347.
  • Aravin, A. and Tusch, T. (2005). FEBS Letters. 579, 5830-5840.
  • Pfeffer, S. et al. (2005). Nat. Meth.. 2, 269-276.
FAQs
Tech Tips

  • Since pre-adenylated oligos are used in the reaction, the ligation efficiency is very high. Usually 1-2 hr at 25C is sufficient to achieve desirable results.

Quality Control Assay
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Specifications
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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
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