Abstract
Based on the standard phosphoramidites chemistry protocol, two oligonucleotides synthetic routes were studied by contact stamping reactants to a modified glass slide. Route A was a contact coupling reaction, in which a nucleoside monomer was transferred and coupled to reactive groups (OH) on a substrate by spreading the nucleoside activated with tetrazole on a polydimethylsiloxane (PDMS) stamp. Route B was a contact detritylation, in which one nucleoside was fixed on the desired synthesis regions where dimethoxytrityl (DMT) protecting groups on the 5’-hydroxyl of the support-bound nucleoside were removed by stamping trichloroacetic acid (TCA) distributed on features on a PDMS stamp. Experiments showed that the synthetic yield and the reaction speed of route A were higher than those of route B. It was shown that 20 mer oligonucleotide arrays immobilized on the glass slide were successfully synthesized using the PDMS stamps, and the coupling efficiency showed no difference between the PDMS stamping and the conventional synthesis methods.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Lu Zuhong, The sfùdy, development, and application of bio-chips, Electronic Technological Report, 1998, 11: 1–5.
Gerhold, D., Rushmore, T., Caskey, C. T., DNA chips: Promising toys have become powerful tools, Techniques, 1993, 3: 168–173.
Khan, J., Bitter, N. L., Chen, Y. D. et al., DNA microarray technology: The anticipated impact on the study of human diease, Biochemica et Biophysca Acta, 1999, 1423: 17–28.
Fodor, S. P. A., Read, J. L., Pirruny, M. C. et al., Light-directed, spatially addressable parallel chemical synthesis, Science, 1991, 251: 767–773.
Schena, M., DNA microarrays, NewYork: Oxford University Press, 1999, 85–97.
Ma Liren, Jiang Zonghua, Bio-chips, Beijing: Chemistry Industry Publishing Company, 2000, 25–47.
Lu Zuhong, A method of preparation of compound micoarray chips and the compound microarray chips produced according to said method, International Patent, WO99/50770, 1998–04–03.
Xia, Y. N., Whitesides, G. M., Soft lithography, Angew Chem. Int. Ed., 1998, 37: 550–575.
Uhlmann, E., Peyman, A., Antisense oligonucleotides: a new therapeutic principle, Chemical Reviews, 1990, 90(4): 544–584.
Weiler, J., Hoeisel, J. D., Combining the preparation of oligonucleotide arrays and synthesis of high-quality primers, Analytical Biochemistry, 1996, 243: 218–227.
Li Jiong, Wang Hong, Zhao Yujie et al., A kind of versatile linkers for covalent bonding DNA on glass surface, in International Forum on Biochip Technologies (ed. Biochip Research & Development Center), Beijing: Tsinghua University, 2000, 492–494.
Xiao Pengfeng, He Nongyue, Lu Zuhong, DNA microarray synthesis using PDMS molecular stamps, I. The fixation and shrinkage of PDMS molecular stamps, Journal of Southeast University (Natural Science Edition), 2001, 2: 6–10.
Septak, M., Kinetic studies on depurination and detritylation of CPG-bound intermediates during oligonucleotide synthesis, Nucleic Acids Research, 1996, 24(15): 3053–3058.
Paul, C. H., Timothy, A. R., Acid binding and detritylation during oligonucleotide synthesis, Nucleic Acids Research, 1996, 24(15): 3048–3052.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Xiao, P., He, N., He, Q. et al. DNA microarray synthesis by using PDMS molecular stamp (II). Sc. China Ser. B-Chem. 44, 442–448 (2001). https://doi.org/10.1007/BF02879820
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF02879820