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Synthesis of hybrid polynucleotide molecules using single-stranded polynucleotide molecules

机译:使用单链多核苷酸分子合成杂合多核苷酸分子

摘要

A method to make libraries of hybrid polynucleotide molecules of two parental polynucleotide molecules utilizing single-stranded DNA was invented. Example of the method comprises several steps: (i) preparation of two single-stranded polynucleotide molecules comprising sequences containing one or more parts of homology and one or more parts of heterology, (ii) random or non-random fragmentation of said polynucleotides, (iii) hybridization of the fragmented molecules followed by de novo polynucleotide synthesis (i.e. polynucleotide chain elongation) on the hybridized molecules, (iv) separation of the chain elongation products (i.e. double-stranded polynucleotide molecules) into single-stranded polynucleotide molecules (denaturation) (v) hybridization of the resultant single-stranded polynucleotide molecules followed by de novo polynucleotide synthesis on the hybridized molecules, and (vi) repeating at least two further cycles of steps (iv) and (v).
机译:发明了一种利用单链DNA制备两个亲本多核苷酸分子的杂合多核苷酸分子的文库的方法。该方法的实例包括几个步骤:(i)制备两个单链多核苷酸分子,其包含含有一个或多个部分同源性和一个或多个部分异源性的序列,(ii)所述多核苷酸的随机或非随机片段化,( iii)片段化的分子杂交,然后在杂交的分子上从头进行多核苷酸合成(即多核苷酸链延长),(iv)将链延长产物(即双链多核苷酸分子)分离成单链多核苷酸分子(变性) (v)杂交所得的单链多核苷酸分子,然后在杂交的分子上从头合成多核苷酸,和(vi)重复步骤(iv)和(v)的至少两个另外的循环。

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