Type of Document Master's Thesis Author Khan, Ahmed Mirza Author's Email Address mahmedk@hotmail.com URN etd-05082008-163131 Title Cyclic Enzymatic Solid Phase Synthesis of DNA Oligonucleotides on an Epoxide-Activated Resin Degree Master of Science Department Chemistry Advisory Committee
Advisor Name Title Markus W. Germann Committee Chair Kathryn B. Grant Committee Member W. David Wilson Committee Member Keywords
- Cyclic enzymatic DNA synthesis
- DNA conjugation to resin
- Labeled DNA oligonucleotide
- Solid phase synthesis
- Epoxide-activated resin
Date of Defense 2008-04-10 Availability unrestricted Abstract Standard chemical DNA synthesis with isotope labels requires expensive reagents; moreover, a large excess of phosphoramadites (typically 50-100 fold) must be used. We developed a process where enzymatic cyclic solid phase synthesis of DNA allows for more economic reagent use. A DNA template was immobilized on an epoxy-activated solid support. This chemistry was chosen because the formed linkage is inert to high pH conditions. High efficiency of the covalent attachment was observed when the reaction was carried out in MgCl2/CAPS buffer. It was found that Mg2+ enables the reaction to be completed over a period of 14 h, compared to 72 h under standard conditions. DNA synthesis was carried in a cyclic fashion on a support bound DNA using Klenow fragment.Files
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