52 research outputs found
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Enhancing Terminal Deoxynucleotidyl Transferase Activity on Substrates with 3' Terminal Structures for Enzymatic De Novo DNA Synthesis.
Enzymatic oligonucleotide synthesis methods based on the template-independent polymerase terminal deoxynucleotidyl transferase (TdT) promise to enable the de novo synthesis of long oligonucleotides under mild, aqueous conditions. Intermediates with a 3' terminal structure (hairpins) will inevitably arise during synthesis, but TdT has poor activity on these structured substrates, limiting its usefulness for oligonucleotide synthesis. Here, we described two parallel efforts to improve the activity of TdT on hairpins: (1) optimization of the concentrations of the divalent cation cofactors and (2) engineering TdT for enhanced thermostability, enabling reactions at elevated temperatures. By combining both of these improvements, we obtained a ~10-fold increase in the elongation rate of a guanine-cytosine hairpin
Enhancing Terminal Deoxynucleotidyl Transferase Activity on Substrates with 3′ Terminal Structures for Enzymatic De Novo DNA Synthesis
Enzymatic oligonucleotide synthesis methods based on the template-independent
polymerase terminal deoxynucleotidyl transferase (TdT) promise to enable the de novo synthesis of
long oligonucleotides under mild, aqueous conditions. Intermediates with a 30
terminal structure
(hairpins) will inevitably arise during synthesis, but TdT has poor activity on these structured
substrates, limiting its usefulness for oligonucleotide synthesis. Here, we described two parallel
efforts to improve the activity of TdT on hairpins: (1) optimization of the concentrations of the
divalent cation cofactors and (2) engineering TdT for enhanced thermostability, enabling reactions at
elevated temperatures. By combining both of these improvements, we obtained a ~10-fold increase in
the elongation rate of a guanine-cytosine hairpin
De novo DNA synthesis using polymerase-nucleotide conjugates
A novel method for de novo DNA synthesis that is based on polymerase-nucleotide conjugates is presented
4,6,8(14)-Trien-3-on-Steroide als fluoreszierende Komponenten in 4,6-Dien-3-on-Steroidpräparaten
Fluorescence of 3-keto-steroids in aqueous solution. Probes for steroid-protein interactions.
The binding of fluorescent 4,6,8(14)-triene-3-one steroids to cyclodextrins as a model for steroid-protein interactions.
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