申请人:President and Fellows of Harvard College
公开号:US10870872B2
公开(公告)日:2020-12-22
The disclosure provides methods for making a polynucleotide wherein the addition of nucleotides can be physically, chemically and/or enzymatically controlled. The methods include combining a selected nucleotide, cations, an error prone or template independent DNA polymerase at a reaction site including an initiator sequence attached thereto and having a 3′ terminal nucleotide, wherein the reaction reagents can be modulated and under conditions that allow covalent addition of one or more of a selected nucleotide to the 3′ terminal nucleotide such that the selected nucleotide becomes a 3′ terminal nucleotide, and repeating the addition step until the polynucleotide is formed.
本公开提供了制造多核苷酸的方法,其中核苷酸的添加可以物理、化学和/或酶法控制。这些方法包括在反应位点上结合所选核苷酸、阳离子、易出错或与模板无关的 DNA 聚合酶,该反应位点包括连接其上并具有 3′末端核苷酸的启动子序列、其中,可以调节反应试剂,并在允许将一种或多种选定核苷酸共价添加到 3′末端核苷酸的条件下,使选定核苷酸成为 3′末端核苷酸,并重复添加步骤,直到形成多核苷酸。