Sequential addition of short DNA oligos in DNA-polymerase-based synthesis reactions
    2.
    发明申请
    Sequential addition of short DNA oligos in DNA-polymerase-based synthesis reactions 有权
    在基于DNA聚合酶的合成反应中连续添加短DNA寡核苷酸

    公开(公告)号:US20050272042A1

    公开(公告)日:2005-12-08

    申请号:US10727779

    申请日:2003-12-03

    IPC分类号: C07H21/04 C12P19/34 C12Q1/68

    摘要: A method of fabricating a DNA molecule of user-defined sequence. The method comprises the steps of preselecting a multiplicity of DNA sequence segments that will comprise the DNA molecule of user-defined sequence, separating the DNA sequence segments temporally, and combining the multiplicity of DNA sequence segments with at least one polymerase enzyme wherein the multiplicity of DNA sequence segments join to produce the DNA molecule of user-defined sequence. Sequence segments may be of length n, where n is an even or odd integer. In one embodiment the length of desired hybridizing overlap is specified by the user and the sequences and the protocol for combining them are guided by computational (bioinformatics) predictions. In one embodiment sequence segments are combined from multiple reading frames to span the same region of a sequence, so that multiple desired hybridizations may occur with different overlap lengths. In one embodiment starting sequence fragments are of different lengths, n, n+1, n+2, etc.

    摘要翻译: 制备用户定义序列的DNA分子的方法。 该方法包括以下步骤:预先选择将包含用户定义序列的DNA分子的多个DNA序列片段,以及将多个DNA序列片段与至少一种聚合酶结合, DNA序列片段连接以产生用户定义序列的DNA分子。 序列段可以是长度为n,其中n是偶或奇整数。 在一个实施例中,期望的杂交重叠的长度由用户指定,并且用于组合它们的序列和协议由计算(生物信息学)预测指导。 在一个实施例中,序列片段从多个读取框架组合以跨越序列的相同区域,使得可以以不同的重叠长度发生多个期望的杂交。 在一个实施方案中,起始序列片段具有不同长度,n,n + 1,n + 2等