DNA microarray synthesis by using PDMS molecular stamp (II)Oligonucleotide on-chip synthesis using PDMS stamp

被引:0
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作者
Pengfeng Xiao
Nongyue He
Quanguo He
Chunxiu Zhang
Yiwen Wang
Zuhong Lu
Jiqing Xu
机构
[1] Southeast University,Key Laboratory for Molecular and Biomolecular Electronics of Ministry of Education
[2] Nanjing E-life Gene Medicine Ltd.,undefined
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关键词
PDMS stamps; contact coupling; contact detritylation; oligonucleotide synthesis;
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学科分类号
摘要
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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页码:442 / 448
页数:6
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