Poly (A) Polymerase

Poly (A) Polymerase

Source: Escherichia coli

Poly (A) Polymerase is E. coli derived polymerase, polyadenylating 3’ termini of RNA.

Description:

• Adds poly (A) tails to the 3'-hydroxyl terminus of RNA (1).

• Ideal for determining molar concentration of polyadenylated RNA (2).

• Suitable for 3'-end labeling of RNA (3).

Unit Definition: One unit is the amount of enzyme required to incorporate 1 nmol of AMP into acid-insoluble form in 10 min at 37°C using tRNA as a substrate.

Storage Conditions: Store at –20°C.

Storage Buffer: 25 mM Tris-HCl (pH 7.9 at 22C), 0.1 mM dithiothreitol, 0.01% (v/v) Triton™X-100, 1 mM EDTA, 0.5 M NaCl and 50% (v/v) glycerol.

Assay Conditions: 50 mM Tris-HCl (pH 7.9 at 22°C), 10 mM MgCl₂, 2.5 mM MnCl₂, 0.25 M NaCl, 0.25 mg/ml tRNA, 0.5 mg/ml bovine serum albumin and 0.1 mM [ α-³²P]ATP. Incubation is at 37°C for 10 min in a reaction volume of 100 µl.

Quality Control: All preparations are assayed for contaminating ribonuclease activity.

References:

1. Sippel, A.E. (1973) Eur. J. Biochem. 37, 31-40.

2. Beltz, W.R. and Ashton, S.H. (1982) Fed. Proc. 41, 1450.

3. Krug, M.S. and Berger, S.L. (1986) Anal. Biochem. 153, 315-323.