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In situ synthesis of oligonucleotide arrays by using soft lithography
| Content Provider | Semantic Scholar |
|---|---|
| Author | Xiao, Pengfeng He, Nongyao Liu, Zhengchun He, Quanguo Sun, Xiao Ting Lu, Zuhong |
| Copyright Year | 2002 |
| Abstract | This paper describes the in situ synthesis of oligonucleotide arrays on glass surfaces by using soft lithography. In this method, based on the standard phosphoramidite chemistry protocol, the coupling was achieved by the glass slide being printed with a set of polydimethylsiloxane (PDMS) microstamps, on which was spread nucleoside monomer and tetrazole mixed solution. The elastic characteristic of PDMS allowed it to make conformal contact with the glass slide in the printing coupling. With regard to the efficiency of the printing coupling, the hybridization microscope images of 20-mer oligonucleotide synthesized via the directly drip-dropped coupling and the contact coupling were compared; the fluorescence intensities of the two methods showed no significant differences. The coupling efficiency was also investigated via an end-labelled fluorescence nucleotide method and a stepwise yield of 97% was obtained. A high-quality, high spatial resolution and large-scale PDMS stamp, which was developed by integrating 168 different microstamps on one glass substrate for synthesizing oligonucleotide arrays. The stamp was modified to improve the surface wettability by plasma discharge treatments, so that microstamps could be used to fabricate oligonucleotide arrays. A motional printing head was developed to improve the contact effect of the glass slide with different microstamps. A higher boiling point solvent was used in the printing coupling to inhibit solvent volatilization and to maintain the consistency of reagents on different features of the microstamp. An effective method was used to eliminate residual reactive nucleosides on chips with small molecules containing a hydroxyl group. A specific oligonucleotide array of four probes both matched and mismatched with the target sequence was fabricated to identify the perfect match and mismatch sequences. |
| Starting Page | 756 |
| Ending Page | 762 |
| Page Count | 7 |
| File Format | PDF HTM / HTML |
| DOI | 10.1088/0957-4484/13/6/312 |
| Volume Number | 13 |
| Alternate Webpage(s) | http://diyhpl.us/~bryan/papers2/bibliographies/papers/Xiao%20et%20al%20-%202002%20-%20In%20situ%20synthesis%20of%20oligonucleotide%20arrays%20by%20usi.pdf |
| Alternate Webpage(s) | http://diyhpl.us/~bryan/papers2/bio/In%20situ%20synthesis%20of%20oligonucleotide%20arrays%20by%20using%20soft%20lithography.pdf |
| Alternate Webpage(s) | https://doi.org/10.1088/0957-4484%2F13%2F6%2F312 |
| Language | English |
| Access Restriction | Open |
| Content Type | Text |
| Resource Type | Article |
