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Home > Products > RNA Reagents > RNA Synthesis > E. coli Poly(A) Polymerase E. coli Poly(A) Polymerase Catalog # Size Concentration Price Qty   M0276L 500 units 5,000 units/ml $252.00 M0276S 100 units 5,000 units/ml $63.00 Prices are in US dollars and valid only for US orders. Download: MSDS PDF For the 3' labeling of RNA For preparing a priming site for cDNA synthesis using oligo-dT Enhances translation of RNA transferred into eukaryotic cells Description: Poly(A) Polymerase catalyzes the template independent addition of AMP from ATP to the 3´ end of RNA. Source: An E. coli strain that carries the cloned Poly(A) Polymerase gene from E. coli (1). Applications: Labeling of RNA with ATP or cordycepin Poly(A) tailing of RNA for cloning or affinity purification Enhances translation of RNA transferred into eukaryotic cells. Reagents Supplied: Poly(A) PolymeraseReaction Buffer (10X) ATP (10 mM) Reaction & Storage Conditions Reaction Conditions: 1X Poly(A) PolymeraseReaction Buffer Supplemented with 1 mM ATP Incubate at 37°C. 1X Poly(A) PolymeraseReaction Buffer: 50 mM Tris-HCl 250 mM NaCl 10 mM MgCl2 pH 7.9 @ 25°C Unit Definition: One unit is defined as the amount of enzyme that will incorporate 1 nmol of AMP into RNA in a 20 µl volume in 10 minutes at 37°C.  Unit Assay Conditions: 1X Poly(A) Polymerase Reaction Buffer, 1 mM rATP and 500 ng 5´ FAM labeled poly A 20-mer RNA in a 20 µl reaction. After incubation at 37°C for 10 minutes the length of the poly(A) addition is determined either by gel electrophoresis or with an automated capillary DNA sequencer. In this assay 5 units of enzyme add approximatley 60 to 80 adenosines to the RNA primer. In these conditons 20 units of enzyme will deplete the rATP. Concentration: 5,000 units/ml Storage Conditions: 20 mM Tris-HCl 300 mM NaCl 1 mM DTT 1 mM EDTA 50% Glycerol 0.1% Triton X-100 Storage Temperature: -20°C Notes General notes: rATP is not included in the buffer supplied. Quality Control for Current Lot Quality control values for a specific lot can be found on the datacard which accompanies each product. Quality Assurance Statement: Poly(A) Polymerase contains no detectable DNAses, RNAses and phosphatases. The purified protein contains no detectable DNA or RNA as determined by ethidium staining of an agarose gel. RNase Assay: Incubation of a 10 µl reaction containing 5 units of E. coli Poly(A) Polymerase with 40 ng of RNA transcript for 2 hours at 37°C resulted in no detectable degradation of the RNA as determined by gel electrophoresis. DNA Exonuclease Activity: Incubation of a 50 µl reaction containing 10 units of Poly(A) Polymerase with 1 µg of a mixture of single and double-stranded 3H E.coli DNA (200,000 cpm/µg) for 3 hours at 37°C released < 0.1% of the total radioactivity. DNA Endonuclease Activity: Incubation of a 10 µl reaction containing 5 units of E. coli Poly(A) Polymerase with 40 ng of RNA transcript for 2 hours at 37°C resulted in no detectable degradation of the RNA as determined by gel electrophoresis. References Cao, G.J. and Sarkar, N. (1992) Proc. Natl. Acad. Sci. USA, 89, 10380-10384. New England Biolabs, Inc. is an ISO 9001 and ISO 14001 Certified Company. NEB certifies that it is a small business in accordance with the US Small Business Administration and 13 CFR 121.201