T4 RNA Ligase

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]rA_14–20 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]rA_14–20.