【文章內(nèi)容簡(jiǎn)介】 address various sequences for high throughput applications, in areas ranging from medicine to environmental analysis. Our new approach in this field is based on an in situ photosynthesis platform that uses digital photolithography to activate photogenerated reactions in a microfluidic chip to produce large amounts of DNA’s or peptides using inexpensive, conventional DNA/peptide building blocks. This technology enables us to gain control over each individual reaction at each reactive site。 to eliminate the usage of photomaskguided photolithography – thus, reducing the cost considerably。 yet maintaining flexibility in design, and feasibility for massively parallel in situ synthesis in our threedimensional chambers on microfluidic chips . CONCLUSION Microarrays have emerged as a useful tool to understand and correlate gene sequences and their related functions。 to extend our knowledge to the proteomic level and to utilize it to capture more information. Our technology enables us to manufacture flexible, highperformance and cost effective microarrays of high fidelity, based on our platform that bines massively parallel lightgated in situ synthesis process, digital photolithography and microfluidic technology. Combined with the above technological advantages, OligoArray , our oligonucleotide design software and OligoArrayDb, our database of predesigned oligonucleotides, allows expression analysis at the genome scale for any anism for which the genome sequence is known, without relying on cDNA or oligonucleotide libraries. REFERENCES: 1. Gao X. et al, ‘A Flexible lightdirected DNA Chip Synthesis Gated By Deprotection Using Solution Photogenerated Acids’, Nucleic Acids Research, 29, 47444750 (2022). 2. Gao X. et al, ‘Oligonucleotide Synthesis Using Solution Photogenerated Acids’, Journal of American Chemical Society, 120, 126989 (1998). 3. Komolpis K. et al, ‘ Lightdirected Simultaneous of Oligopeptides on Microarray Substrate Using a Photogenerated Acid’, Biotechnology Progress, 18, 641646 (2022). 4. Leproust E. et al, ‘Digital LightDirected Synthesis: A Microarray Platform That Permits Rapid Reaction Optimization on a Combinatorial Basis’, Journal of Combinatorial Chemistry, 2, 349354 (2022). 5. Pellois J. P. et al, ‘ Individually Addressable Parallel Peptide Synthesis on Microchips’, Nature Biotechnology, 20, 922926 (2022). 6. Rouillard J. M. et al, ‘OligoArray: GenomeScale Oligonucleotide Design for Microarrays’, Bioinformatics, 18, 4867 (2022). ACKNOWLEDGEMENTS: DARPA ATO MICHIGAN LIFE SCIENCES CORRIDOR FUND GRANT NO. 1805 COLLABORATIONS: MICHIGAN STATE UNIVERSITY UNIVERSITY OF HOUSTON UNIVERSITY OF MICHIGANANN ARBOR MEDICAL SCHOOL XEOTRON CORPORATION OLIGONUCLEOTIDE DESIGN FOR DNA MICROARRAYS: A THERMODYNAMIC APPROACH Our technology of DNA synthesis on chips bined with the availability of an increasing number of sequenced genomes, have prompted us to implement a software to design specific oligonucleotides for microarrays. Ideally, such oligonucleotides should be totally specific to their respective targets to avoid any crosshybridization and should not form stable secondary structures that may interfere with the labeled probes during hybridization. We have developed OligoArray (Rouillard et al, 2022), a program to design specific oligonucleotides at genome scale. The latest version, OligoArray (Rouillard et al. submitted), uses a thermodynamic approach to predict secondary structures and calculate the specificity of a probe on the chips to a unique target in a mixture of labeled targets. PANGENOMIC DATABASE OF OLIGONUCLEOTIDES We have used our OligoArray software to design pangenomic sets of oligonucleotides devoted to gene expression analysis for most of the anisms with a sequenced genome. As of today, we have designed 774,642 oligonucleotides representing 277,976 transcripts from 80 anisms (14 archaea, 59 bacteria and 7 eucaryotes including Homo sapiens and some geic models). We have successfully designed at least one oligonucleotide for more than 99% of all transcripts from all anisms processed and there is at least one specific oligonucleotide for 94% of these transcripts. All these oligonucleotide sets are stored into OligoArrayDb, a public database that allows not only to retrieve a plete set of oligonucleotides, but also to build subsets according to user defined keywords (Rouillard et al, submitted). Supplementary information: IN SITU SYNTHESIS The binatorial synthesis has brought about novel opportunities for efficiently creating reactions of a scaffold chemical moiety with a number of different pounds to yield a library of various pounds with different substituent groups, . nucleotides, amino acids. A digital microarray platform consisting of a microarray reactor, a micromirror array image projector, a controller puter and an automated synthesizer. By in situ synthesis, an array of oligonucleotides is synthesized using conventional DMTprotected phosphoramidite chemistry. This is different from conventional synthesis by the use of a photogenerated acid (PGA) rather than an acid in DMT deprotection step to control the parallel synthesis. Deprotection is initiated by directing light at selected threedimensional chambers in microfluidic chips. Upon activation, acid forms in seconds, removing DMT group. Parallel synthesis using acidlabile protecting groups. The DNA chain is deprotected in a spatially controlled manner using photogenerated acid (PGA) (b and d), followed by coupling and oxidation reactions (c and e). This cycle is repeated until the desired lengths and sequences are obtained (f). hv hv hv hv T