Supplementary data: An improved gene synthesis method with asymmetric directions of oligonucleotides designed based on a simulation program

Table S1 Name, length and nucleotide sequence of oligonucleotide used in AESOE reaction

Table S2 Integer-number pairs simulating annealing and extension reaction of ssDNA

Figure S1 Gel electrophoresis of AESOE-synthesized DNA-fragments Five μl of the AESOE reaction solution taken out after the cycle shown on the photograph was analyzed by 1.5% agarose electrophoresis. M: Size marker (100, 200, 300, 400 and 500 bases from the bottom). The name of the oligomer set is shown on the left of the photograph.

Figure S3 Schematic diagram of the incomplete dimer from which an extension reaction is initiated. The direction of the arrow indicates the direction of the base sequence of the oligomer (5’->3’).

Figure S4 Schematic diagram of the dimers from which any extension reaction is not initiated. The direction of the arrow indicates the direction of the base sequence of the oligomer (5’->3’).

Figure S5 Schematic diagram of tendency on reconstruction of original duplex of DNA comparing incomplete dimer formation. The direction of the arrow indicates the direction of the base sequence of the oligomer (5’->3’). Black and red arrows indicate original duplex DNA and short oligomer respectively.

Figure S6 Schematic diagram of tendency on dimer formation. Possibility of hybridization with short oligomers is higher than with long oligomers. The direction of the arrow indicates the direction of the base sequence of the oligomer (5’->3’).

Figure S7 Schematic diagram of tendency on dimer formation. Possibility of hybridization with oligomers present in high concentration is higher than those in low concentration. The direction of the arrow indicates the direction of the base sequence of the oligomer (5’->3’).

Supplement methods. Simulator for Oligomer Polymerization