Our artificial gene synthesis process starts with oligonucleotide synthesis. Multiple long oligos of around 50-60 bases in length with similar melting temperature are chemically synthesized as fragments based on the desired final gene sequence. Oligos are designed to assemble with each other through overlapping sequences.
Next the gene is assembled by using PCR. The oligo fragments are assembled into blocks of up to 1kb of double stranded DNA. These blocks are then assembled and amplified once again using PCR to create one large double stranded DNA construct. The synthesized DNA is inserted into a specific vector. The vector is then isolated and amplified. Next colonies are grown and appropriate ones are then selected containing the synthetic gene of interest.
All final artificial genes are verified by DNA sequencing and only those with the correct sequences are selected. This ensures that the gene synthesized conforms to the specifications planned in the beginning of the project. Further QC measures are applied and the gene is lyophilized into a microcentrifuge tube. The gene is now ready for downstream applications.