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T4 RNA Ligase

Joins ssRNA or ssDNA onto the 5´-Phosphoryl End of ssRNA or ssDNA

  • Adds [5´-32P] nucleoside 3´,5´-bis (phosphate) onto single-stranded RNA
  • Can be inactivated by heating at 65°C for 15 minutes
  • Supplied at a concentration of 10u/μl

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Description

Size: 500u
Catalog number: M1051

T4 RNA Ligase catalyzes the ATP-dependent ligation of single-stranded RNA or DNA onto the 5´-phosphoryl termini of single-stranded RNA or DNA. The enzyme, purified from recombinant E. coli CA4 (RNase I-deficient), has an apparent molecular weight of 43.5kDa. T4 RNA Ligase is provided with 10X Reaction Buffer: 500mM Tris-HCl (pH 7.8 at 25°C), 100mM MgCl2, 50mM DTT, 10mM ATP.

Applications

  • 3´ end-labeling of RNA.
  • Circularization of RNA and DNA oligonucleotides.
  • Cloning full-length cDNA.

References

  1. Silber, R. et al. (1972) Proc. Natl. Acad. Sci. USA 69, 3009–13.
  2. England, T. et al. (1977) Proc. Natl. Acad. Sci. USA 74, 4839–42.
  3. Uhlenbeck, O.C. and Gumport, R.I. (1982) The Enzymes 158, 31.
  4. Brennan, C.A. et al. (1983) Meth. Enzymol. 100, 38–52.
  5. Schaefer, B.C. (1995) Anal. Biochem. 227, 255–73.

Storage Buffer: 10mM Tris (pH 7.5 at 25°C), 50mM KCl, 0.1mM EDTA, 1mM DTT, 50% glycerol, 0.1% Tween® 20.

Source: Recombinant E. coli strain.

QC Tests: Activity, SDS-PAGE/purity, DNase, RNase, endonuclease.

Unit Definition: One unit catalyzes the formation of 1nmol of [5´-32P]rA14–20 into a phosphatase-resistant form in 30 minutes at 37°C in a total volume of 30μl. The reaction conditions are: 50mM Tris-HCl (pH 7.8 at 25°C), 10mM MgCl2, 5mM DTT, 1mM ATP and 10μM [5´-32P]rA14–20.

Protocols

Complete Protocol

T4 RNA Ligase Protocol


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What’s in the box?
Size: 500u

Item Part # Size
T4 RNA Ligase M105A 1 × 500u
T4 RNA Ligase 10X Buffer M107A 1 × 500μl

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

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