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T4 RNA Ligase 1 (ssRNA Ligase)

Catalog # Concentration Size List Price Quantity Your Price
M0204L 10000 units/ml 5000 units $435.00
$391.50
M0204S 10000 units/ml 1000 units $109.00
$98.10
Catalog # Size List Price Your Price
M0204L 5000 units $435.00
$391.50
M0204S 1000 units $109.00
$98.10
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca
T4 RNA Ligase 1 catalyzes the ligation of a 5´ phosphoryl-terminated nucleic acid donor to a 3´ hydroxyl-terminated nucleic acid acceptor through the formation of a 3´ → 5´ phosphodiester bond with hydrolysis of ATP to AMP and PPi. Substrates include single-stranded RNA and DNA as well as dinucleoside pyrophosphates.
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    Avoiding RNase Contamination
 
Product Source
An E. Coli strain that carries the T4 RNA Ligase I gene
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  T4 RNA Ligase 1 (ssRNA Ligase) M0204SVIAL -20 1 x 0.1 ml 10,000 units/ml
  T4 RNA Ligase Reaction Buffer B0216SVIAL -20 1 x 1.5 ml 10 X
  Adenosine-5'-Triphosphate (ATP) B0756AVIAL -20 1 x 0.2 ml 10 mM
  PEG 8000 B1004SVIAL -20 1 x 1 ml 1 X
  T4 RNA Ligase 1 (ssRNA Ligase) M0204LVIAL -20 1 x 0.5 ml 10,000 units/ml
  T4 RNA Ligase Reaction Buffer B0216SVIAL -20 1 x 1.5 ml 10 X
  PEG 8000 B1004SVIAL -20 2 x 1 ml 1 X
  Adenosine 5'-Triphosphate (ATP) P0756SVIAL -20 1 x 1 ml 10 mM
Application Features
  • Labeling of 3'-termini of RNA with 5'-[32P] pCp (3)
  • Inter- and intramolecular joining of RNA and DNA molecules (4,5)
  • Synthesis of single-stranded oligodeoxyribo-nucleotides (6)
  • Incorporation of unnatural amino acids into proteins (7)
  • Ligation of ss-RNA and DNA

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to convert 1 nanomole of 5´-[32P]rA16 into a phosphatase-resistant form in 30 minutes at 37°C.

Reaction Conditions

1X T4 RNA Ligase Reaction Buffer 
Supplement with 1 mM ATP
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
50 mM KCl
1 mM DTT
0.1 mM EDTA
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation

65°C for 15 minutes

Unit Assay Conditions

1X T4 RNA Ligase reaction buffer, supplemented with 1 mM ATP, is mixed with the RNA substrate (10µM of 5´-[32P]rA16 ) and varying amounts of enzyme. Incubation is at 37°C for 15 minutes (8).

Concentration:
10,000 units/ml

Companion Products
Materials Sold Separately
Notes
  • Addition of DMSO to 10% (v/v) is required for pCp ligation (3).
References
  • England, T., Gumport, R. and Uhlenbeck, O. (1977). Proc. Natl. Acad. Sci. USA. 74, 4839-4842.
  • Rand, K.N. and Gait, M.J. (1984). EMBO J.. 3, 397-402.
  • England, T. and Uhlenbeck, O. (1978). Nature. 275, 560-562.
  • Romaniuk, P. and Uhlenbeck, O. (1983). R. Wu, L. Grossman and K. Moldave(Ed.), Methods Enzymol.. 100, 52-56. New York: Academic Press.
  • Moore, M.J. and Sharp, P.A. (1992). Science. 256, 992-997.
  • Tessier, D.C., Brousseau, R., and Vernet, T. (1986). Anal. Biochem.. 158, 171-178.
  • Noren, C.J. et al. (1989). Science. 244, 182-188.
  • Silber, R., Malathi, B.G. and Hurwitz, J. (1972). Proc. Natl. Acad. Sci. USA. 69, 3009-3013.
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

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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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