Fast and efficient chemical ligation of oligonucleotides using diels-alder reaction.
Diels-Alder chemical ligation was applied to the ligation of two and three oligonucleotides to form a single, continuous strand with an unnatural DNA backbone at the ligation site. The reaction occurred between a maleimide-labelled oligonucleotide and a furan-labelled oligonucleotide to form two ty...
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| Format: | Final Year Project |
| Language: | English |
| Published: |
2010
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| Subjects: | |
| Online Access: | http://hdl.handle.net/10356/39857 |
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| Institution: | Nanyang Technological University |
| Language: | English |
| Summary: | Diels-Alder chemical ligation was applied to the ligation of two and three oligonucleotides to form a single, continuous strand with an unnatural DNA backbone at the ligation site. The reaction occurred between a maleimide-labelled oligonucleotide and a furan-labelled
oligonucleotide to form two types of Diels-Alder linkage (one of each directionality). A
different type of Diels-Alder ligated oligonucleotide was formed via the reaction between dual-furan labelled oligonucleotide and two maleimide-labelled oligonucleotide. The template-mediated reaction occurred rapidly and efficiently, with most product formation occurring by zero hour reaction time. An approximately drop in 10 °C was found for the melting temperature of oligonucleotides with a single unnatural backbone as compared to native DNA strand,
indicative of distortion of the duplex by the linker. |
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