.rte-hr { background: linear-gradient(to right, white, #162F65); border: 0; height: 1px; } .bio-chart { border: solid #3361ac 2px; text-align: center; margin-bottom: 15px; border-radius: 12px; box-shadow: 0px 0px 5px rgba(22, 47, 101, 0.5); } .bio-chart-title { background-color: #162f65; color: white; display: flex; padding: 5px 0; border-radius: 9px 9px 0 0; } .bio-chart-row:nth-child(odd) { background-color: #ebf6ff; display: flex; padding: 5px 0; } .bio-chart-row:nth-child(even) { background-color: white; display: flex; padding: 5px 0; } .bio-chart-row:last-child { border-radius: 0 0 12px 12px; } .chart-col-8 { width: 66.67%; } .chart-col-4 { width: 33.33%; } .chart-component { color: #3361ac; font-weight: bold; } .chart-parameter { color: #162f65; } .operation-recommend-icon { margin-top: 3px; } .operation-alert { background-color: #ffecd9; border-radius: 8px; padding: 10px; margin: 0 5px; display: flex; gap: 10px; color: #c62828; box-shadow: 0 0 5px rgba(22, 47, 101, 0.2); margin-bottom: 20px; } .operation-alert-icon { margin-top: 3px; } Product Information 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.coliConcentration: 5U/μLUnit 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. Key points of operation Protocol: The standard reaction will produce ～150bp long poly(A)-tail on up to 10μg 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. Combine the following reagents: Component Amount RNA 1-10μg 10X poly(A) Polymerase Reaction Buffer 2μL ATP (10mM) 2μL Poly(A) Polymerase 20 units (Optional) RNase Inhibitor 25μL Nuclease-free H2O to 20μL Incubate reaction at 37°C for 60 minutes. 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. Security Information Storage Conditions: -20℃Quality Assurance: Free of endonuclease, exonuclease and RNase activities.Physical Purity: ＞95% by SDS-PAGE. Reference 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.
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