Poly(A) Polymerase Tailing Kit
- Addition of a poly(A) tail to RNA synthesized in vitro in order to increase the stability of the RNA and enhance its ability to be translated in vivo after transfection or microinjection into eukaryotic cells.1-3
- Addition of a poly(A) tail to an RNA molecule or a mixture of RNA molecules in order to provide a priming site for synthesis of first-strand cDNA using a primer with poly(dT) on its 3´ end.
- Synthesis of polyadenylated RNA for nucleic acid amplification methods or gene expression studies.
- 3´-end labeling of RNA with radioactive A residues.4
- Quantifying mRNA.5
- High enzymatic purity, specificity, and activity.
- Easy, rapid, and highly efficient poly(A) tailing of RNA (Fig. 1).
Storage Buffer: 50% glycerol containing 50 mM Tris-HCl (pH 7.5), 100 mM NaCl, 1 mM DTT, 0.1 mM EDTA, and 0.1% Triton® X-100.
10X Reaction Buffer: 0.5 M Tris-HCl (pH 8.0), 2.5 M NaCl, and 100 mM MgCl2. A separate 10-mM ATP Solution is also provided.
Quality Control: Poly(A) Polymerase is tested for polyadenylation of RNA in vitro. It is free of detectable exo- and endonuclease and RNase activity.
- Drummond, D.R. et al. (1985) J. Cell. Biol. 100, 1148.
- Galili, G. et al. (1988) J. Biol. Chem. 263, 5764.
- Belasco, J. and Brawerman, G. (1993) Control of Messenger RNA Stability, Academic Press, San Diego, CA.
- Lingner, J. and Keller, W. (1993) Nucleic Acids Res. 21, 2917.
- Krug, M.S. and Berger, S.L. (1987) Methods Enzymol. 152, 262.
- Gething, M. et al. (1980) Nature 287, 301.
||Figure 1. Poly(A) tailing of RNA with the Poly(A) Polymerase Tailing Kit. The entire final product of an AmpliCap-Max™ in vitro transcription capping reaction (60 µg of a 1,760-base luciferase RNA) was treated with 8 units of Poly(A) Polymerase for the indicated times. For each sample, 0.1-µg aliquots were analyzed on a 1% agarose-formaldehyde gel.|