An Improved Gene Synthesis Method with Asymmetric Directions of Oligonucleotides Designed using a Simulation Program

Abstract

Artificial gene synthesis based on oligonucleotide augmentation is known as overlap extension PCR which generates a variety of intermediate synthetic products. The orientation and concentration of oligomers can be adjusted to reduce the synthesis of intermediates and optimize the full-length process of DNA synthesis, using a simulation program for serial oligomer extension. The efficiency of the serial oligomer extension process is predicted to be greatest when oligomers are in a ‘forward-reverse-reverse-reverse’ direction. Oligomers with such designed directions demonstrated generation of the desired product in the shortest time (number of cycles) by repeated annealing and elongation. This method, named Asymmetric Extension supported by a Simulator for Oligonucleotide Extension (AESOE), has shown efficiency and effectiveness with potentials for future improvements and optimal usage in DNA synthesis.

Designer DNA biomolecules as a defined biomaterial for 3D bioprinting applications

“…Three different approaches are currently utilized for the synthesis of DNA, such as DNA replication, 153 polymerase chain reaction (PCR), 154 and artificial gene synthesis. 155 In most organisms, DNA replication happens using the existing DNA as a template except for some retrovirus, in which the RNA is the genetic material. Several DNA replication models, such as semi-conservative replication 156 and rolling circle amplification (RCA) 157 have explained about the replication mechanisms in prokaryotes and eukaryotes.…”