Description
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 MgCl 2 , 50mM DTT, 10mM ATP.
Applications
3´ end-labeling of RNA.
Circularization of RNA and DNA oligonucleotides.
Cloning full-length cDNA.
References
Silber, R. et al. (1972) Proc. Natl. Acad. Sci. USA 69 , 3009–13.
England, T. et al. (1977) Proc. Natl. Acad. Sci. USA 74 , 4839–42.
Uhlenbeck, O.C. and Gumport, R.I. (1982) The Enzymes 158 , 31.
Brennan, C.A. et al. (1983) Meth. Enzymol. 100 , 38–52.
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´- 32 P]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 MgCl 2 , 5mM DTT, 1mM ATP and 10μM [5´- 32 P]rA 14–20 .
