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Enzymatic synthesis and modification of high molecular weight DNA using terminal deoxynucleotidyl transferase. 2019

Sonal Deshpande, and Yunqi Yang, and Ashutosh Chilkoti, and Stefan Zauscher
Department of Biomedical Engineering, Duke University, Durham, NC, United States.

The recognition that nucleic acids can be used as polymeric materials led to the blossoming of the field of DNA nanotechnology, with a broad range of applications in biotechnology, biosensors, diagnostics, and drug delivery. These applications require efficient methods to synthesize and chemically modify high molecular weight DNA. Here, we discuss terminal deoxynucleotidyl transferase (TdT)-catalyzed enzymatic polymerization (TcEP) as an alternative to conventional enzymatic and solid-phase DNA synthesis. We describe biochemical requirements for TcEP and provide step-by-step protocols to carry out TcEP in solution and from surfaces.

UI MeSH Term Description Entries
D008970 Molecular Weight The sum of the weight of all the atoms in a molecule. Molecular Weights,Weight, Molecular,Weights, Molecular
D004247 DNA A deoxyribonucleotide polymer that is the primary genetic material of all cells. Eukaryotic and prokaryotic organisms normally contain DNA in a double-stranded state, yet several important biological processes transiently involve single-stranded regions. DNA, which consists of a polysugar-phosphate backbone possessing projections of purines (adenine and guanine) and pyrimidines (thymine and cytosine), forms a double helix that is held together by hydrogen bonds between these purines and pyrimidines (adenine to thymine and guanine to cytosine). DNA, Double-Stranded,Deoxyribonucleic Acid,ds-DNA,DNA, Double Stranded,Double-Stranded DNA,ds DNA
D004253 DNA Nucleotidylexotransferase A non-template-directed DNA polymerase normally found in vertebrate thymus and bone marrow. It catalyzes the elongation of oligo- or polydeoxynucleotide chains and is widely used as a tool in the differential diagnosis of acute leukemias in man. EC 2.7.7.31. Terminal Addition Enzyme,Terminal Deoxyribonucleotidyltransferase,Deoxynucleotidyl Transferase,Deoxynucleotidyltransferase,Desoxynucleotidyl Transferase,Desoxynucleotidyltransferase,Tdt Antigen,Terminal Deoxynucleotidyl Transferase,Terminal Deoxyribonucleotidyl Transferase,Addition Enzyme, Terminal,Antigen, Tdt,Deoxynucleotidyl Transferase, Terminal,Deoxyribonucleotidyl Transferase, Terminal,Deoxyribonucleotidyltransferase, Terminal,Enzyme, Terminal Addition,Nucleotidylexotransferase, DNA,Transferase, Deoxynucleotidyl,Transferase, Desoxynucleotidyl,Transferase, Terminal Deoxynucleotidyl,Transferase, Terminal Deoxyribonucleotidyl
D058105 Polymerization Chemical reaction in which monomeric components are combined to form POLYMERS (e.g., POLYMETHYLMETHACRYLATE). Polymerizations

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