Modern Approaches to de novo Synthesis of Extended DNA Fragments: Assembly of a Wide Repertoire of Sequences

T A Semashko; G Y Fisunov; E A Tsoy; D R Kharrasov; I K Chudinov; D V Evsyutina; G Y Shevelev; V M Govorun

doi:10.32607/actanaturae.27362

Modern Approaches to de novo Synthesis of Extended DNA Fragments: Assembly of a Wide Repertoire of Sequences

Acta Naturae. 2024 Jan-Mar;16(1):77-85. doi: 10.32607/actanaturae.27362.

Authors

T A Semashko^{1

2}, G Y Fisunov^{1

2}, E A Tsoy^{1

2}, D R Kharrasov¹, I K Chudinov¹, D V Evsyutina^{1

2}, G Y Shevelev³, V M Govorun¹

Affiliations

¹ Research Institute for Systems Biology and Medicine, Moscow, 117246 Russian Federation.
² Lopukhin Federal Research and Clinical Center of Physical-Chemical Medicine, Moscow, 119435 Russian Federation.
³ Institute of Chemical Biology and Fundamental Medicine, Novosibirsk, 630090 Russian Federation.

Abstract

The standardization of DNA fragment assembly methods for many tasks of synthetic biology is crucial. This is necessary for synthesizing a wider repertoire of sequences, as well as for further automation and miniaturization of such reactions. In this work, we proposed conditions for the assembly of DNA fragments from chemically synthesized oligonucleotides and we identified the errors occurring in the sequence under these conditions. Additionally, we proposed conditions for further combining synthetic fragments into larger DNA fragments. We showed that the optimized conditions are suitable for the assembly of a wide range of sequences.

Keywords: DNA artificial synthesis; DNA synthesis; PCR assembly; gene assembly; gene synthesis; polymerase cyclic assembly; spermidine.