title: Enzymatic polymerisation involving 2'-amino-LNA nucleotides.
abstract: The triphosphate of the thymine derivative of 2'-amino-LNA (2'-amino-LNA-TTP) was synthesised and found to be a good substrate for Phusion® HF DNA polymerase, allowing enzymatic synthesis of modified DNA encoded by an unmodified template. To complement this, 2'-amino-LNA-T phosphoramidites were incorporated into DNA oligonucleotides which were used as templates for enzymatic synthesis of unmodified DNA using either KOD, KOD XL or Phusion polymerases. 2'-Amino-LNA-T in the template and 2'-amino-LNA-TTP as a substrate both decreased reaction rate and yield compared to unmodified DNA, especially for sequences with multiple 2'-amino-LNA-T nucleotides.
authors.affiliation: Nucleic Acid Center, Department of Physics, Chemistry and Pharmacy, University of Southern Denmark, Odense M, Denmark.
authors.forename: Marie W
authors.initials: MW
authors.lastname: Johannsen
authors.forename: Rakesh N
authors.initials: RN
authors.lastname: Veedu
authors.forename: Andreas Stahl
authors.initials: AS
authors.lastname: Madsen
authors.forename: Jesper
authors.initials: J
authors.lastname: Wengel
mesh_terms: D001483:Base Sequence; D001704:Biopolymers / Q000378:metabolism*; D017931:DNA Primers; D004798:Enzymes / Q000378:metabolism*; D009841:Oligonucleotides / Q000378:metabolism*
chemical_list: D001704:Biopolymers; D017931:DNA Primers; D004798:Enzymes; D009841:Oligonucleotides; C477371:locked nucleic acid