Poly(A) Polymerase (1000 U)

This QIAGEN enzyme catalyzes the addition of AMP from ATP to the 3ʹ hydroxyl of RNA, a reaction that requires Mg²⁺ and is template independent. It is intended for making poly(A)-tailed RNA and for 3ʹ end RNA labeling. The protein is produced by an E. coli strain expressing the Poly(A) Polymerase gene from a plasmid.

Quality control specifications include a purity of >95% and a specific activity of >20,000 U/mg. Additional release tests report: single-stranded exonuclease <5% released (200 U); double-stranded exonuclease <1% released (200 U); double-stranded endonuclease no conversion (200 U); E. coli DNA contamination <10 copies (100 U); and no detectable non-specific RNase contamination (200 U).

The enzyme is supplied in 25 mM Tris-HCl, 500 mM NaCl, 1 mM MgCl₂, 0.1 mM DTT, 0.1 mM EDTA and 50% glycerol at pH 8.0 (25°C). The accompanying 10x reaction buffer contains 500 mM Tris-HCl, 2.5 M NaCl, and 100 mM MgCl₂ at pH 7.9 (25°C). One unit is defined as the amount of enzyme that incorporates 1 nmol of ATP into acid-insoluble material in 10 minutes at 37°C. A standard reaction is assembled in a total volume of 20 µL and incubated at 37°C for 30 minutes; the reaction can be stopped by adding EDTA to a final concentration of 10 mM or proceeding to a cleanup step.