Enzymatic polymerisation involving 2′-amino-LNA nucleotides

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.

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