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Protocols for Poly(U) Polymerase FAQs for RNA Reagents Technical Reference for RNA Reagents Enzyme Finder NEBcutter NEBuffer Chart Double Digest Finder PCR Selection Tool Isoschizomers DNA Sequences and Maps REBASE NEBuffer 2 Murine RNase Inhibitor RNase Inhibitor

Home > Products > RNA Reagents > RNA Synthesis > Poly(U) Polymerase Poly(U) Polymerase Catalog # Size Concentration Price Qty   M0337S 60 units 2,000 units/ml $75.00 Prices are in US dollars and valid only for US orders. Download: MSDS PDF Description: Poly(U) Polymerase catalyzes the template independent addition of UMP from UTP or AMP from ATP to the 3´ end of RNA. Source: An E. coli strain that carries the cloned poly(U) polymerase gene of Schizosaccharomyces pombe Cid1. Applications: Labeling of RNA with UTP Poly(U) tailing of RNA for cloning Studying effects of poly(U) tailing on stability and translation of RNA transferred into eukaryotic cells Poly (A) tailing of 2' O-Me modified 3' ends Reagents Supplied: NEBuffer 2 (10X) Reaction & Storage Conditions Reaction Conditions: 1X NEBuffer 2 Supplemented with 1 mM UTP Incubate at 37°C. 1X NEBuffer 2: 10 mM Tris-HCl 50 mM NaCl 10 mM MgCl2 1 mM Dithiothreitol pH 7.9 @ 25°C Unit Definition: One unit is defined as the amount of enzyme that incorporates 1 nmol of UMP into RNA in a 50 µl volume in 10 minutes at 37°C.  Unit Assay Conditions: 1X NEBuffer 2, 0.5 mM 3H UTP and 5 µg yeast RNA are combined in a 50 µl reaction incubated at 37°C for 10 minutes. Concentration: 2,000 units/ml Storage Conditions: 10 mM Tris-HCl 100 mM NaCl 1 mM DTT 0.1 mM EDTA 50% Glycerol Storage Temperature: -20°C Notes General notes: UTP is not included in the buffer. Poly(U) Polymerase in NEBuffer 2 will incorporate rNMP from rNTP into RNA. Tailing length of poly(U) varies with UTP. Poly(U) Polymerase is highly processive under low primer concentrations (<100 pmol). Protocols Protocols for Poly(U) Polymerase 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(U) Polymerase contains no detectable DNAses, and RNAses. The purified protein contains no detectable DNA or RNA. RNase Assay:: Incubation of a 10 µl reaction containing 2 units of Poly(U) Polymerase with 40 ng of RNA transcript for 16 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(U) Polymerase with 1 cg 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 50 µl reaction containing 10 units of Poly(U) Polymerase with 1 µg of supercoiled plasmid for 4 hours at 37°C resulted in <10% conversion to nicked molecules as determined by agarose gel electrophoresis. References Wickens, M. and Kwak, J.E.  (2008) Science, 319, 1344. Kwak, J.E. and Wickens, M. ( (2008) RNA, 13, 860. Rissland, O.O.S., Mikulasova, A. and Norbury, C.J.  (2007) Molecular and Cell Biology, 27, 3612. Reagents Sold Separately NEBuffer 2 Companion Products Murine RNase Inhibitor RNase Inhibitor 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