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Measuring DNA polymerising enzyme activity in body fluids, cells or virus infected cultures

机译:测量体液,细胞或病毒感染培养物中的DNA聚合酶活性

摘要

Method for determining the activity of a nucleotide-polymerising enzyme in a sample comprises: (a) capturing the enzyme with a monoclonal antibody (MAb) immobilised on a fixed carrier (the MAb binds the enzyme without significantly affecting its activity); (b) removing impurities and/or disturbance factors; (c) starting nucleotide polymerisation by adding a reaction soln. contg. a primer/template construction and nucleotide substrate; (d) providing reaction conditions to favour permanent association between the enzyme and primer/template prod. complex; (e) washing the nucleotide substrate, primer/template and reaction soln. to remove freshly synthesised polymer; (f) measuring the amt. of nucleotide incorporated with the polymer by standard techniques, and (g) determining enzyme activity.
机译:用于确定样品中核苷酸聚合酶活性的方法包括:(a)用固定在固定载体上的单克隆抗体(MAb)捕获酶(MAb结合酶而不显着影响其活性); (b)去除杂质和/或干扰因素; (c)通过添加反应溶液开始核苷酸聚合。续引物/模板结构和核苷酸底物; (d)提供反应条件以促进酶与引物/模板产物之间的永久缔合。复杂; (e)洗涤核苷酸底物,引物/模板和反应溶液。去除新鲜合成的聚合物; (f)量度。通过标准技术与聚合物结合的核苷酸的测定,和(g)测定酶活性。

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