E. coli derived polymerase, polyadenylating 3" termini of RNA
Adds poly A tails to the 3'-hydroxyl terminus of RNA. Labels 3'-ends of RNA with a [α-32P] cordycepin triphosphate. Ideal for determining molar concentration of polyadenylated RNA.
50 mM Tris-HCl (pH 7.9).10 mM MgCl2. 2.5 mM MnCl2. 250 mM NaCl. 25 μg tRNA. 0.1 mM [α-32P] ATP. 50 μg bovine serum albumin. Reaction volume of 100 μl.
Incubation at 37 °C
500 Units; 100 Units
25mM Tris-HCl (pH 7.9) 0.1 mM DTT 0.01% (v/v) Triton X-100 1.0 mM EDTA 0.5M NaCl 50% (v/v) glycerol
One unit is the amount of enzyme required to incorporate 1.0 nmol of labeled ATP into acid insoluble form for 10 minutes at 37 °C under the stated assay conditions.