DNA microarray synthesis by using PDMS molecular stamp (II) |
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Authors: | Pengfeng Xiao Nongyue He Quanguo He Chunxiu Zhang Yiwen Wang Zuhong Lu Jiqing Xu |
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Institution: | 1. Key Laboratory for Molecular and Biomolecular Electronics of Ministry of Education, Southeast University, 210096, Nanjing, China 2. Nanjing E-life Gene Medicine Ltd., 210016, Nanjing, China
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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. |
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