Method for Generating of Oligonucleotide Arrays Using In Situ Block Synthesis Approach

摘要

The idea of this invention is to prepare ordered oligonucleotides arrays from two or more pre-synthesized shorter parts—block-synthesis approach. The parts are linked together enzymatically to form a full-length oligonucleotide of a desired sequence. Such an approach allows splitting the oligonucleotide sequences into common and unique parts. It gives the possibility to place the functional group on a common part and to minimize the length of the unique parts. Method of the invention allows combinatorial synthesis of position-specific regions. Using combinatorial approach, position-specific regions are generated by linking two or more unique oligonucleotides, so that just few said unique oligonucleotides give rise to a large variety of codes, for example, 10 unique oligonucleotides linked pairwise can produce 100 position-specific regions. In comparison to preparation of oligonucleotide arrays by spotting of full-length sequences, suggested approach is more cost-efficient, allows flexibility in generating position-specific unique sequences and is less prone to oligonucleotide length restrictions. In comparison to in situ synthesis of oligonucleotides from nucleotides, current invention allows cost-efficient solution for synthesis of oligonucleotides with free 3′ ends. Important application of the current invention is preparation of two-dimensional oligonucleotide arrays for preparation of sequencing libraries from 2D distributed NA molecules. Oligonucleotides on such arrays need to have position-specific sequences and free 3′ ends for further enzymatic reactions.