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T4 RNA Ligase 1 (ssRNA Ligase), High Concentration

Catalog # Concentration Size List Price Quantity Your Price
M0437M 30000 units/ml 5000 units $435.00
$391.50
Catalog # Size List Price Your Price
M0437M 5000 units $435.00
$391.50
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 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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T4 RNA Ligase 1, High Concentration (M0437M), is a 3X higher concentration than our standard T4 RNA Ligase 1 (M0204S/L). This product is packaged with a vial of 100 mM ATP (N0451A). The increased concentration of reaction components is helpful in lower volume reactions where reagent volumes are limited.

T4 RNA Ligase 1 catalyzes the ligation of a 5´ phosphoryl-terminated nucleic acid donor to a 3´ hydroxyl-terminated nucleic acceptor through the formation of a 3´ → 5´ phosphodiester bond, with hydrolysis of ATP to AMP and PPi. Substrates include singlestranded RNA and DNA as well as dinucleoside pyrophosphates (1).
Product Source
An E. coli strain that carries the T4 RNA Ligase 1 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), High Concentration M0437MVIAL -20 1 x 0.167 ml 30,000 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
  ATP N0437AVIAL -20 1 x 0.1 ml 100 mM
Application Features
  • Ligation of ss-RNA and DNA
  • 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 oligodeoxyribonucleotides (6)
  • Incorporation of unnatural amino acids into proteins (7)

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

50 mM KCl
10 mM Tris-HCl
0.1 mM EDTA
1 mM DTT
50% Glycerol
pH 7.5 @ 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).

Companion Products
Notes
  • Addition of DMSO to 10% (v/v) isrequired 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). In R. Wu, L. Grossman and K. Moldave(Ed.), Methods in Enzymology. Vol. 100, (pp. 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.
Citations
Additional Citations
  • Jesse M. Engreitz, Klara Sirokman, Patrick McDonel, Alexander A. Shishkin, Christine Surka, Pamela Russell, Sharon R. Grossman, Amy Y. Chow, Mitchell Guttman, and Eric S. Lander (2014) RNA-RNA Interactions Enable Specific Targeting of Noncoding RNAs to Nascent Pre-mRNAs and Chromatin Sites Cell 159(1), 188-99.PubMedID: 25259926, DOI: 10.1016/j.cell.2014.08.018
  • Engreitz JM, Pandya-Jones A, McDonel P, Shishkin A, Sirokman K, Surka C, Kadri S, Xing J, Goren A, Lander ES, Plath K, and Guttman M (2013) The Xist lncRNA exploits three-dimensional genome architecture to spread across the X chromosome Science 341(6147), 1237973.PubMedID: 23828888, DOI: 10.1126/science.1237973
  • Guttman M, Russell P, Ingolia NT, Weissman JS, Lander ES (2013) Ribosome profiling provides evidence that large noncoding RNAs do not encode proteins Cell 154(1), 240-51.PubMedID: 23810193, DOI: 10.1016/j.cell.2013.06.009
Publications
  • Shishkin AA, Giannoukos G, Kucukural A, Ciulla D, Busby M, Surka C, Chen J, Bhattacharyya RP, Rudy RF, Patel MM, Novod N, Hung DT, Gnirke A, Garber M, Guttman M, Livny J (2015). Simultaneous generation of many RNA-seq libraries in a single reaction Nat�Methods.PubMedID: 25730492, DOI: 10.1038/nmeth.3313
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
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