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Assembly Fabrication of Oligonucleotide Arrays

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Abstract:

In this paper, a simple, reliable and flexible method for fabricating oligonucleotide arrays integrating in in-situ synthesis with a spotting technique is described. In this approach, different oligonucleotide sequences were synthesized on coded modification glass slides using combinatorial chemistry and a mature phosphoramidite chemistry protocol. The slides were then sliced into smaller pieces. Finally, an oligonucleotide array was fabricated by arbitrarily assembling the different coded pieces onto another solid support. A 5 × 5 array including four different sequences of the P16 gene and a control (blank) was successfully assembled. The results indicated that the hybridization fluorescence intensities from the same sequences located at different places on the array were homogeneous and uniform. Background fluorescence was much lower. The fluorescence intensity ratio of a matched sequence to a one-base mismatched sequence, a two-base mismatched sequence and a three-base mismatched sequence was 0.499, 0.236 and 0.04, respectively. The results for the same sequence at different spots in the chip were reproducible with the relative standard deviation ranging from 6.64% to 10.2% (n = 5). This method has the advantages of high probe-density of in-situ synthesis, and off-chip flexibility. Moreover, it does not need any immobilization process to bond oligonucleotides on the substrate.

Keywords: ASSEMBLY FABRICATION; COMBINATIONAL SYNTHESIS; OLIGONUCLEOTIDE ARRAY

Document Type: Research Article

DOI: https://doi.org/10.1166/jnn.2005.205

Publication date: 2005-08-01

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  • Journal for Nanoscience and Nanotechnology (JNN) is an international and multidisciplinary peer-reviewed journal with a wide-ranging coverage, consolidating research activities in all areas of nanoscience and nanotechnology into a single and unique reference source. JNN is the first cross-disciplinary journal to publish original full research articles, rapid communications of important new scientific and technological findings, timely state-of-the-art reviews with author's photo and short biography, and current research news encompassing the fundamental and applied research in all disciplines of science, engineering and medicine.
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