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Method of Making Polynucleotides Using Closed-Loop Verification

机译:闭环验证法制备多核苷酸的方法

摘要

A method for making a polynucleotide is provided including (a) delivering one or more reaction reagents including an error prone or template independent DNA polymerase, cations and a selected nucleotide to a reaction site including an initiator sequence having a terminal nucleotide for a time period and under conditions capable of covalently adding one or more of the selected nucleotide to the terminal nucleotide at the 3′ end of the initiator such that the selected nucleotide becomes a terminal nucleotide, and (b) determining whether the selected nucleotide has been added to the terminal nucleotide, wherein if the selected nucleotide has not been added to the terminal nucleotide, then repeating step (a) until the selected nucleotide has been added, and (c) repeating steps (a) and (b) until the polynucleotide is formed.
机译:提供了一种制备多核苷酸的方法,该方法包括:(a)在一段时间内将包含易于出错或不依赖模板的DNA聚合酶,阳离子和选择的核苷酸的一种或多种反应试剂递送至包括具有末端核苷酸的引发剂序列的反应位点,以及在能够将一种或多种选择的核苷酸共价添加到引发剂的3'端的末端核苷酸的条件下,使得选择的核苷酸成为末端核苷酸,并且(b)确定选择的核苷酸是否已经被添加到末端核苷酸,其中如果尚未将选择的核苷酸添加到末端核苷酸,则重复步骤(a)直到选择的核苷酸已添加,以及(c)重复步骤(a)和(b)直到形成多核苷酸。

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