Poly(A) Polymerase

Poly(A) Polymerase
Catalog No.
>95% by SDS-PAGE
Recombinant E.coli

Product description: Poly(A) Polymerase uses ATP as a substrate for template-independent addition of adenosine monophosphate to the 3'-hydroxyl termini of RNA molecules.

Source: Recombinant E.coli

Concentration and Size: 5U/μL

Unit Definition: One unit is defined as the amount of enzyme that will incorporate 1nmol of AMP into RNA in a 20μL volume in 10 minutes at 37°C.

Storage Buffer: 20mM Tris-HCl (pH7.5@25℃), 1mM EDTA, 300mM NaCl, 1mM DTT, 0.1%(v/v) Triton X-100,50%(v/v) glycerol.

Companion Product: 10X poly(A) Polymerase Reaction Buffer, Cat#ON-127, 500mM Tris-HCl, (pH7.9@25℃), 2.5M NaCl, 100mM MgCl2.

Protocol: The standard reaction will produce ~150bp long poly(A)-tail on up to 10ug of capped or uncapped RNA from IVT reactions and co-transcriptional capping reactions need to be purified before adding to the poly(A)-tailing reaction.

1. Combine the following reagents:

2. Incubate reaction at 37°C for 60 minutes.

3. Stop reaction by adding EDTA to final concentration of >11mM.

* Before poly(A)-tail reaction, heat-denaturation of the RNA may improve the efficiency of adding poly(A)-tail.

Storage Conditions: -20℃

Quality Assurance: Free of endonuclease, exonuclease and RNase activities.

Physical Purity: >95% by SDS-PAGE.

1. Bernstein P and Ross J (1989) Poly(A), poly(A) binding protein and the regulation of mRNA stability. Trends Biochem Sci 14: 373–377.

2. Gallie DR (1991) The cap and poly(A) tail function synergistically to regulate mRNA translational efficiency. Genes Dev 5: 2108–2116.

3. Krug, M.S. and Berger, S.L. (1987) Methods Enzymol. 152, 262.